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Methods 195 (2021) 15–22
Available online 25 May 2021
1046-2023/© 2021 Elsevier Inc. All rights reserved.
Test positivity – Evaluation of a new metric to assess epidemic dispersal
mediated by non-symptomatic cases
Folorunso O. Fasina a,*
, Mudasiru A. Salami b
, Modupe M. Fasina c
, Olutosin A. Otekunrin d
,
Almira L Hoogesteijn e
, James B. Hittner f
a
Food and Agriculture Organization of the United Nations, Dar es Salam, Tanzania & Department of Veterinary Tropical Diseases, University of Pretoria, South Africa
b
College of Medicine, University College Hospital, University of Ibadan, Ibadan, Nigeria
c
Potchefstroom Hospital, North West, South Africa
d
Department of Agricultural Economics and Farm Management, Federal University of Agriculture, Abeokuta (FUNAAB), Nigeria
e
Human Ecology, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Mérida, Mexico
f
Department of Psychology, College of Charleston, Charleston, SC, USA
A R T I C L E I N F O
Keywords:
COVID-19
Test positivity
Geo-epidemiology
Resource-limited countries
Infection
A B S T R A C T
Epidemic control may be hampered when the percentage of asymptomatic cases is high. Seeking remedies for this
problem, test positivity was explored between the first 60 to 90 epidemic days in six countries that reported their
first COVID-19 case between February and March 2020: Argentina, Bolivia, Chile, Cuba, Mexico, and Uruguay.
Test positivity (TP) is the percentage of test-positive individuals reported on a given day out of all individuals
tested the same day. To generate both country-specific and multi-country information, this study was imple­
mented in two stages. First, the epidemiologic data of the country infected last (Uruguay) were analyzed. If at
least one TP-related analysis yielded a statistically significant relationship, later assessments would investigate
the six countries. The Uruguayan data indicated (i) a positive correlation between daily TP and daily new cases
(r = 0.75); (ii) a negative correlation between TP and the number of tests conducted per million inhabitants
(TPMI, r = − 0.66); and (iii) three temporal stages, which differed from one another in both TP and TPMI me­
dians (p < 0.01) and, together, revealed a negative relationship between TPMI and TP. No significant rela­
tionship was found between TP and the number of active or recovered patients. The six countries showed a
positive correlation between TP and the number of deaths/million inhabitants (DMI, r = 0.65, p < 0.01). With
one exception –a country where isolation was not pursued− , all countries showed a negative correlation between
TP and TPMI (r = 0.74). The temporal analysis of country-specific policies revealed four patterns, characterized
by: (1) low TPMI and high DMI, (2) high TPMI and low DMI; (3) an intermediate pattern, and (4) high TPMI and
high DMI. Findings support the hypothesis that test positivity may guide epidemiologic policy-making, provided
that policy-related factors are considered and high-resolution geographical data are utilized.
1. Introduction
Unlike most other infections, COVID-19 is an infection characterized
by a high percentage of asymptomatic cases: at least 40% and up to 80%
of the cases may not reveal symptoms [1–3]. Such a feature creates an
unprecedented problem; when this disease disseminates, in principle,
the only way to identify all infected individuals would be through a
universal testing program, i.e., testing, repeatedly, 100% of the popu­
lation. At least in 2020, no country achieved such a high level of testing:
on average, countries tested much less than 1% of the population, on a
given day. Given the scarcity of testing resources, in 2020, Austria and
Germany reported that not more than 20% of the cases were likely to be
detected [4].
Consequently, in 2020, the true epidemiologic status of the popula­
tion was unknown. To remedy such a difficult and global problem,
which may be encountered, again, in future epidemics caused by this,
similar or other infections, new metrics may be required.
Testing populations seems to differ markedly from testing in­
dividuals. While clinical medical practices have historically emphasized
personalized diagnostics, population-oriented testing is needed when
epidemics are rapidly disseminating [5,6].
Test positivity is a metric of potential relevance in situations in which
* Corresponding author.
E-mail address: daydupe2003@yahoo.co.uk (F.O. Fasina).
Contents lists available at ScienceDirect
Methods
journal homepage: www.elsevier.com/locate/ymeth
https://doi.org/10.1016/j.ymeth.2021.05.017
Received 20 December 2020; Received in revised form 21 May 2021; Accepted 23 May 2021
Methods 195 (2021) 15–22
16
the percentage of asymptomatic cases is high. Test positivity (TP) is the
percentage of test-positive individuals reported on a given day out of all
individuals tested the same day. The importance of TP in the context of
the COVID-19 pandemic was first mentioned on March 30, 2020: when
the World Health Organization described this metric in a public con­
ference [7]. While now broadly recognized, TP has been empirically
examined. Hence, TP, as well as associated variables (including time),
should be explicitly evaluated.
The ‘test positivity rate’ (TP) or ‘percent positive rate’ may help
public health officials answer questions such as: ‘Are we doing enough
testing for the amount of people who are getting infected?’ [8]. When the
total number of tests conducted is too low, the test positivity (TP) per­
centage may be high. A high TP percentage may indicate that there are
more people with coronavirus in the community who have not yet been
tested [9]. A high TP percentage may reflect a rapid epidemic dissemi­
nation and, consequently, it may predict that places with a high TP will
soon express a high or very high number of deaths/million inhabitants
[6]. Vice versa, a low TP percentage may prognosticate the imminent
cessation of an outbreak.
Other indicators that may influence TP or be influenced by TP are: a)
the number of tests conducted per million inhabitants (TPMI), b) the
number of deaths reported per million inhabitants (DPMI), c) the
number of active cases (all test-positive cases minus deaths and recov­
ered patients), and d) the number of recovered patients.
However, no variable may demonstrate utility or validity unless it is
explored within an explicit –not an assumed–geographical and temporal
context. That is so because the connectivity that explains epidemic
dispersal is a geo-temporal structure [10–12].
Fig. 1. Epidemic curve and early metrics for COVID-19 infections, Uruguay, March to May, 2020. (A). Daily test positivity against epidemic days; (B). Daily number
of new cases versus daily test positivity; (C). Cumulative cases and deaths over the period, March to May 2020; (D). Cases and tests per million inhabitants; (E).
Fatalities and tests per million inhabitants; (F). Deaths per million inhabitants and tests per million inhabitants.
There was a positive correlation between the daily number of new cases versus daily test positivity (r= 0.75 [CI= 0.61, 0.85]), Fig. 1B.
F.O. Fasina et al.
Methods 195 (2021) 15–22
17
Consequently, here the percentage of test positivity was investigated
in Argentina, Bolivia, Chile, Cuba, Mexico and Uruguay –a group of
countries selected because the first COVID-19 case, in these countries,
was reported less than six weeks apart. Therefore, a relatively similar
temporal timeframe allowed a hypothesis-generating study that
considered both specific time points and geography. The purpose of this
study was to generate the first geo-temporal evaluation of several
epidemiologic variables associated with test positivity.
2. Methods
Daily epidemiologic reports corresponding to Uruguay were
collected from the Uruguayan National System of Emergencies [13].
Because the data source was published in Spanish, it was corroborated
with daily reports made by Worldometer [14]. Data corresponding to
Argentina, Bolivia, Chile, Cuba, Mexico and Uruguay were collected
from OurWorldInData [15]. Temporal data from these countries on test
positivity, TPMI and DPMI are reported in Supplementary Table 1.
The period under study was the first 60–90 epidemic days, which
correspond (for the group of six countries) to February-May 2020.
Specifically, daily and/or cumulative count data on: (i) the number of
tests conducted in a given day, (ii) the number of test-positive in­
dividuals reported in a given day, (iii) the number of active cases, and
(v) number of deaths were obtained. A commercial package (Minitab 19,
State College, PA, Minitab Inc.) was used to perform statistical analyses
and create plots. Population medians were analyzed using the non-
parametric Mann-Whitney test.
3. Results
The data corresponding to Uruguay were investigated first. The
rationale for such a priority was the fact that it was the last country
affected by this pandemic within the selected group and, consequently,
epidemic patterns observed in Uruguay could follow and/or mimic
patterns also exhibited by other countries prior to March 13, 2020
(when the first case was detected in Uruguay). The second reason for this
research plan was that Uruguay started to develop diagnostic tests
before the first case was detected and, consequently, the analysis of test
positivity could be facilitated [16].
The first estimates of test positivity were rather high in Uruguay.
However, after approximately two weeks, they decreased and remained
low in the first 60 epidemic days (Fig. 1A). TP was positively correlated
with the daily number of new cases (r = 0.75, Fig. 1B).
An alternative way to monitor the progression of this pandemic is to
plot the cumulative number of test-positive individuals (cases) together
with the cumulative number of SARS-CoV2-positive fatalities. This
approach revealed two curves, which were not parallel (Fig. 1C).
Therefore, fatalities could not be predicted from the case data.
A similar finding was revealed when the cumulative number of cases
was compared to the cumulative number of tests performed per million
inhabitants (TPMI): the curves were not parallel (Fig. 1D). One data
inflection revealed that, between two and three epidemic weeks
(after ~ 2000 TPMI were reached), the number of TPMI increased lin­
early over time (Fig. 1D). A similar pattern was observed when the cu­
mulative number of fatalities was compared to TPMI: after TPMI
reached 1000, the number of deaths flattened out (Fig. 1E). These
Fig. 2. Inferential statistics on COVID-19, Uruguay, March to May, 2020. (A). Daily test positivity against epidemic days and tests per million inhabitants; (B). Daily
test positivity versus cumulative tests per million inhabitants reveal an L-shaped curve; (C). Temporal evaluation of daily test positivity reveals three distinct stage
over the period, March to May 2020; (D). Cumulative cases and cumulative active cases plotted against a temporal timeline.
There was a negative correlation between the daily test positivity and tests/million inhabitants (r= - 0.66 [CI= - 0.79, - 0.48]), Fig. 2A.
F.O. Fasina et al.
Methods 195 (2021) 15–22
18
patterns were also noticed when TPMI was compared to the cumulative
number of deaths reported per million inhabitants (DPMI): only after
TPMI exceeded 1000, the DPMI values flattened out (Fig. 1F).
Several of these variables showed at least two phases (Fig. 1 C, E, F).
The earliest stage displayed a quasi-exponential growth (i.e., the number
of cases grew faster than time, which resulted in a quasi-vertical
pattern). Later (after a data inflection was revealed) a quasi-linear
phase was observed, in which the growth in the number of cases was
slower than time and, therefore, cases exhibited a quasi-horizontal
pattern (e.g., Fig. 1C).
Test positivity was negatively correlated with testing in Uruguay
(r = − 0.66, Fig. 2A). Three subsets of temporal observations showed
decreasing values of TP, which corresponded to increasing (and statis­
tically significantly different) values of TPMI (p < 0.01, Fig. 2B, C).
When the sub-group of active cases was compared to all cases (which
included deaths and recovered cases), the cumulative number of active
cases exhibited a data inflection followed by a decreasing trend which,
together, supported the view that patterns displayed by the data on
active cases may inform earlier than the alternative indicator (Fig. 2D).
Test positivity was positively correlated with deaths/million in­
habitants (DPMI) when the six countries were simultaneously investi­
gated (r = 0.65, Fig. 3A). However, when the same countries were
investigated in terms of TPMI and TP, a distinct, non-linear pattern was
detected (Fig. 3B). When one seemingly outlier country was removed
from the analysis, a negative correlation was revealed (r = − 0.74,
Fig. 3C). When DPMI and TPMI were investigated, a non-linear (L-sha­
ped) distribution was noticed (Fig. 3D). When time, TP, and DPMI were
plotted, three patterns were detected, which were characterized by a)
low TP and low DPMI (a pattern displayed by three countries), b) in­
termediate values (shown by two countries), and c) high TP and high
DPMI, which were only revealed by one country (Fig. 3E). In contrast,
when TPMI was plotted together with time and TP, four patterns were
Fig. 3. External validity and comparison of observed patterns for COVID-19 in six countries. (A). Deaths per million inhabitants against test positivity; (B). Tests per
million inhabitants against test positivity; (C). Tests per million inhabitants against test positivity, excluding Chile; (D). Deaths per million inhabitants against tests
per million inhabitants, excluding Chile; (E). Three-dimensional plot of deaths per million inhabitants against temporal scale and test positivity; (F). Three-
dimensional plot of tests per million inhabitants against temporal scale and test positivity.
There was a positive correlation for five countries except Chile, between the test positivity and deaths per million inhabitants (r= 0.65 [CI= 0.44, 0.80]), Fig. 3A. However,
there was a negative correlation between test positivity and tests per million inhabitants after excluding Chile (r= - 0.74 [CI= - 0.86, - 0.55]). This demonstrates that TP may be
a reproducible metric, provided that the context and other variables are also considered.
F.O. Fasina et al.
Methods 195 (2021) 15–22
19
observed, which revealed a) high TPMI and low TP values (found in two
countries), b) a lower TPMI but a higher TP than the previous subset (a
profile shown in one country), c) the lowest TPMI and highest TP per­
centages (detected in two countries), and d) high TPMI and high TP
values (a pattern observed in one country, Fig. 3F).
When geography was considered, the association between TPMI and
DPMI was further documented (Fig. 4A-F). Within two weeks, one
country partially predicted the fatalities associated with epidemic
dispersal: the lowest TPMI values preceded an ~250% growth in DPMI.
However, exceptions to this pattern were also noticed: in the same
period of time, two countries that also displayed low TPMI values
showed a negligible change in DPMI (Fig. 4A-F).
The temporal fluctuation of TP values was minimal across countries.
In the period under study and without exceptions, countries showed
either one- or two-digit percentages (Fig. 5A-F).
In addition to TP-related geo-temporal assessments, the shape of the
data on active cases was also informative. For instance, the detection of
data inflections could be viewed as a positive outcome (Fig. 6A–C).
Similarly, both the directionality and numerical values of the ratio be­
tween recovered and active cases could be considered: when the ratio is
greater than 1, a positive prognosis may be considered. In addition,
when time progresses faster than the epidemiologic metric (when a
horizontal directionality predominates) a rather stagnant situation may
be inferred. In contrast, when the ratio of recovered individuals over all
active cases grows faster than time (when a vertical directionality is
noticed), a positive prognosis is defensible (Fig. 7A-C).
4. Discussion
Hoping to explore a novel metric –test positivity− , we evaluated
Fig. 4. Country-level comparison of cases, mortality and test positivity per million inhabitants for COVID-19. Plot of deaths per million inhabitants against tests per
million inhabitants in (A). April 24, 2020; (B). April 27, 2020; (C). April 28, 2020; (D). May 4, 2020; (E). May 6, 2020; (F). May 9, 2020.
Fig. 4 (A – F) show a synopsis of a geo-temporal analysis; it includes (low-resolution or aggregate) geographical data and a temporal description of epidemics that started less
than 6 weeks apart. For visual comparisons, each plot is divided into 4 quadrants (’low & low’, ’high & low’, ’low & high’, and ’high & high’) It is shown that high TP is
associated with a faster growth of deaths/mill inh. The opposite pattern (low test positivity & low deaths/mil inh) shows one exception.
F.O. Fasina et al.
Methods 195 (2021) 15–22
20
several epidemic parameters associated with SARS CoV-2 for the first 60
to 90 epidemic days in six countries where the time when the first
COVID-19 case was reported differed in six or fewer weeks. Given that
test positivity is not a direct measure of health status at the population
level, we wondered whether this metric could estimate –albeit indi­
rectly–the progression (or lack) of epidemic dispersal.
The answer to such a question is unlikely to be totally elucidated
with a single study. However, one study may provide information that
supports or rejects theories and/or technologies aiming at monitoring
population health status when exposed to infectious diseases with a high
or very high percentage of asymptomatic individuals.
Earlier studies have shown that test positivity predicts epidemic
dispersal better than the simple number of tests performed per million
inhabitants [5,6]. In addition, this evaluation documented several sta­
tistically significant associations that may reflect biological relation­
ships. For instance, when two sources of data were used, the Uruguayan
case revealed a negative correlation between TP and TPMI, both when it
was investigated alone and when four other countries were explored
(Figs. 2A and 3C). Such a reproducibility (which involves two data
sources and five countries) does not support random findings.
The fact that several variables also exhibited reproducible patterns
(such as two temporal phases), provides a group of metrics associated
with TP that may be worth considering in future studies. Some of these
indicators may provide geography- (country-), and time-specific infor­
mation potentially useful in epidemic monitoring. For instance, in the
Uruguayan epidemic, at least 1000 tests per million inhabitants
appeared to be needed to flatten out the growth of fatalities (Fig. 1E). In
all six investigated countries, the simultaneous consideration of TPMI
and DPMI, over time, may help to find both similarities and differences
(Fig. 3E and F).
The observed relationships among TP, tests/million inhabitants, the
shape of active cases, and the shape of the recovered/active cases (R/A)
ratio support the WHO’s recommendations on policies aimed at low TP
percentages: the highest levels of testing (which may detect credible
levels of disease prevalence) only occurred at the lowest levels of TP.
The test positivity, together with the shape of the number of active cases,
can determine whether and when the epidemic is/is not under control.
Associated patterns –such as increasing or decreasing trends of the
recovered/active ratio data–may inform policy makers whether an
overflow of hospital beds and associated resources is/is not likely in the
following weeks. When TP is analyzed together with tests/million in­
habitants, it may indicate (in a country-specific manner and without pre-
Fig. 5. Selected country classification by cases, mortality and test positivity for COVID-19. Plot for (A). Uruguay; (B). Chile; (C). Cuba; (D). Bolivia; (E). Argentina;
(F). Mexico.
Fig. 5 (A – F) show the same data depicted in fig 4, now at individual countries. The left column (A, C, E) displays countries with one digit of test positivity percentages. The right
column (B, D, F) shows two-digit test positivity percentages.
F.O. Fasina et al.
Methods 195 (2021) 15–22
21
established assumptions) whether testing is sufficient or not. While the
data analyzed in this study referred to nationwide levels and aggregated
data tend to be less informative because they may lose resolution [17],
these metrics could also be used with lower-scale, higher-resolution
(county- or city-level) geographical data [11,12].
While limited in scope, this study generated hypotheses. One
example is the hypothesis that lack of explicit isolation policies may
inhibit the effectiveness of massive testing. Such a hypothesis was
illustrated by one country that showed a high level of testing, but did not
pursue isolation [18]. Patterns observed were compatible with studies
that have reported (a) lockdowns but no specific testing scheme, and (b)
neither testing nor lockdowns [18].
Thus, this initial study on test positivity provides support for several
recommendations. We advocate for a policy that focuses on four aspects:
(1) massive and repeated testing, (2) border control, (3) tracing of every
case, and (4) active isolation which, to be successful, must be imple­
mented within a Critical Response Time, that is, before the exponential
growth kicks in [19]. Effective responses are not likely to depend on
piecemeal approaches that only consider one or a few strategies. Instead,
new and accelerated educational programs are needed, which should
provide training on several disciplines, including but not limited to (a)
geography, (b) epidemiology, (c) the economics of public health, (d)
development and use of new software packages that facilitate early (real
time) and geographically specific interventions, and (e) the creation of
new systems that both collect and disseminate data.
Provided that additional studies reproduce these findings, one
possible application of test positivity is to become an alternative to the
Fig. 6. Temporal-related metrics in the analysis of COVID-19 cases in three
countries. (A) Argentina; (B) Cuba; (C). Uruguay.
Fig. 7. Plots of temporal pattern of the ratio between recovered and active
cases in selected countries. (A) Argentina; (B) Cuba; (C). Uruguay.
F.O. Fasina et al.
Methods 195 (2021) 15–22
22
quasi-centennial ‘reproductive number’ (or R0), which estimates the
ratio of secondary over primary cases. That is so because, at the present
time, there are no real time estimates on how rapidly epidemics may
progress and such estimates are also prone to numerous sources of error
[20,21]. Unlike the lack of geographically explicit information and
delayed calculations associated with the R0 (which prevent real time,
site-specific decision-making), test positivity can be calculated on daily
basis and provide actual (not assumed) high-resolution, geographical
information (e.g., neighborhood- or village-specific geographical co­
ordinates). Such an application could improve policy-making: instead of
assuming test positivity is homogeneously distributed over large terri­
tories and remains constant over time, daily changes at specific (and
geographically small) sites could support context-specific, cost-benefit
oriented interventions [6].
To win the war against the next epidemic, reactive responses are not
acceptable. Proactive responses are the only alternative, which require a
robust and immediate creation of novel educational programs of inter/
transdisciplinary nature. To that end, further research on the advantages
and limitations of test positivity is necessary.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://doi.
org/10.1016/j.ymeth.2021.05.017.
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F.O. Fasina et al.

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Test positivity – Evaluation of a new metric to assess epidemic dispersal mediated by non-symptomatic cases

  • 1. Methods 195 (2021) 15–22 Available online 25 May 2021 1046-2023/© 2021 Elsevier Inc. All rights reserved. Test positivity – Evaluation of a new metric to assess epidemic dispersal mediated by non-symptomatic cases Folorunso O. Fasina a,* , Mudasiru A. Salami b , Modupe M. Fasina c , Olutosin A. Otekunrin d , Almira L Hoogesteijn e , James B. Hittner f a Food and Agriculture Organization of the United Nations, Dar es Salam, Tanzania & Department of Veterinary Tropical Diseases, University of Pretoria, South Africa b College of Medicine, University College Hospital, University of Ibadan, Ibadan, Nigeria c Potchefstroom Hospital, North West, South Africa d Department of Agricultural Economics and Farm Management, Federal University of Agriculture, Abeokuta (FUNAAB), Nigeria e Human Ecology, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Mérida, Mexico f Department of Psychology, College of Charleston, Charleston, SC, USA A R T I C L E I N F O Keywords: COVID-19 Test positivity Geo-epidemiology Resource-limited countries Infection A B S T R A C T Epidemic control may be hampered when the percentage of asymptomatic cases is high. Seeking remedies for this problem, test positivity was explored between the first 60 to 90 epidemic days in six countries that reported their first COVID-19 case between February and March 2020: Argentina, Bolivia, Chile, Cuba, Mexico, and Uruguay. Test positivity (TP) is the percentage of test-positive individuals reported on a given day out of all individuals tested the same day. To generate both country-specific and multi-country information, this study was imple­ mented in two stages. First, the epidemiologic data of the country infected last (Uruguay) were analyzed. If at least one TP-related analysis yielded a statistically significant relationship, later assessments would investigate the six countries. The Uruguayan data indicated (i) a positive correlation between daily TP and daily new cases (r = 0.75); (ii) a negative correlation between TP and the number of tests conducted per million inhabitants (TPMI, r = − 0.66); and (iii) three temporal stages, which differed from one another in both TP and TPMI me­ dians (p < 0.01) and, together, revealed a negative relationship between TPMI and TP. No significant rela­ tionship was found between TP and the number of active or recovered patients. The six countries showed a positive correlation between TP and the number of deaths/million inhabitants (DMI, r = 0.65, p < 0.01). With one exception –a country where isolation was not pursued− , all countries showed a negative correlation between TP and TPMI (r = 0.74). The temporal analysis of country-specific policies revealed four patterns, characterized by: (1) low TPMI and high DMI, (2) high TPMI and low DMI; (3) an intermediate pattern, and (4) high TPMI and high DMI. Findings support the hypothesis that test positivity may guide epidemiologic policy-making, provided that policy-related factors are considered and high-resolution geographical data are utilized. 1. Introduction Unlike most other infections, COVID-19 is an infection characterized by a high percentage of asymptomatic cases: at least 40% and up to 80% of the cases may not reveal symptoms [1–3]. Such a feature creates an unprecedented problem; when this disease disseminates, in principle, the only way to identify all infected individuals would be through a universal testing program, i.e., testing, repeatedly, 100% of the popu­ lation. At least in 2020, no country achieved such a high level of testing: on average, countries tested much less than 1% of the population, on a given day. Given the scarcity of testing resources, in 2020, Austria and Germany reported that not more than 20% of the cases were likely to be detected [4]. Consequently, in 2020, the true epidemiologic status of the popula­ tion was unknown. To remedy such a difficult and global problem, which may be encountered, again, in future epidemics caused by this, similar or other infections, new metrics may be required. Testing populations seems to differ markedly from testing in­ dividuals. While clinical medical practices have historically emphasized personalized diagnostics, population-oriented testing is needed when epidemics are rapidly disseminating [5,6]. Test positivity is a metric of potential relevance in situations in which * Corresponding author. E-mail address: daydupe2003@yahoo.co.uk (F.O. Fasina). Contents lists available at ScienceDirect Methods journal homepage: www.elsevier.com/locate/ymeth https://doi.org/10.1016/j.ymeth.2021.05.017 Received 20 December 2020; Received in revised form 21 May 2021; Accepted 23 May 2021
  • 2. Methods 195 (2021) 15–22 16 the percentage of asymptomatic cases is high. Test positivity (TP) is the percentage of test-positive individuals reported on a given day out of all individuals tested the same day. The importance of TP in the context of the COVID-19 pandemic was first mentioned on March 30, 2020: when the World Health Organization described this metric in a public con­ ference [7]. While now broadly recognized, TP has been empirically examined. Hence, TP, as well as associated variables (including time), should be explicitly evaluated. The ‘test positivity rate’ (TP) or ‘percent positive rate’ may help public health officials answer questions such as: ‘Are we doing enough testing for the amount of people who are getting infected?’ [8]. When the total number of tests conducted is too low, the test positivity (TP) per­ centage may be high. A high TP percentage may indicate that there are more people with coronavirus in the community who have not yet been tested [9]. A high TP percentage may reflect a rapid epidemic dissemi­ nation and, consequently, it may predict that places with a high TP will soon express a high or very high number of deaths/million inhabitants [6]. Vice versa, a low TP percentage may prognosticate the imminent cessation of an outbreak. Other indicators that may influence TP or be influenced by TP are: a) the number of tests conducted per million inhabitants (TPMI), b) the number of deaths reported per million inhabitants (DPMI), c) the number of active cases (all test-positive cases minus deaths and recov­ ered patients), and d) the number of recovered patients. However, no variable may demonstrate utility or validity unless it is explored within an explicit –not an assumed–geographical and temporal context. That is so because the connectivity that explains epidemic dispersal is a geo-temporal structure [10–12]. Fig. 1. Epidemic curve and early metrics for COVID-19 infections, Uruguay, March to May, 2020. (A). Daily test positivity against epidemic days; (B). Daily number of new cases versus daily test positivity; (C). Cumulative cases and deaths over the period, March to May 2020; (D). Cases and tests per million inhabitants; (E). Fatalities and tests per million inhabitants; (F). Deaths per million inhabitants and tests per million inhabitants. There was a positive correlation between the daily number of new cases versus daily test positivity (r= 0.75 [CI= 0.61, 0.85]), Fig. 1B. F.O. Fasina et al.
  • 3. Methods 195 (2021) 15–22 17 Consequently, here the percentage of test positivity was investigated in Argentina, Bolivia, Chile, Cuba, Mexico and Uruguay –a group of countries selected because the first COVID-19 case, in these countries, was reported less than six weeks apart. Therefore, a relatively similar temporal timeframe allowed a hypothesis-generating study that considered both specific time points and geography. The purpose of this study was to generate the first geo-temporal evaluation of several epidemiologic variables associated with test positivity. 2. Methods Daily epidemiologic reports corresponding to Uruguay were collected from the Uruguayan National System of Emergencies [13]. Because the data source was published in Spanish, it was corroborated with daily reports made by Worldometer [14]. Data corresponding to Argentina, Bolivia, Chile, Cuba, Mexico and Uruguay were collected from OurWorldInData [15]. Temporal data from these countries on test positivity, TPMI and DPMI are reported in Supplementary Table 1. The period under study was the first 60–90 epidemic days, which correspond (for the group of six countries) to February-May 2020. Specifically, daily and/or cumulative count data on: (i) the number of tests conducted in a given day, (ii) the number of test-positive in­ dividuals reported in a given day, (iii) the number of active cases, and (v) number of deaths were obtained. A commercial package (Minitab 19, State College, PA, Minitab Inc.) was used to perform statistical analyses and create plots. Population medians were analyzed using the non- parametric Mann-Whitney test. 3. Results The data corresponding to Uruguay were investigated first. The rationale for such a priority was the fact that it was the last country affected by this pandemic within the selected group and, consequently, epidemic patterns observed in Uruguay could follow and/or mimic patterns also exhibited by other countries prior to March 13, 2020 (when the first case was detected in Uruguay). The second reason for this research plan was that Uruguay started to develop diagnostic tests before the first case was detected and, consequently, the analysis of test positivity could be facilitated [16]. The first estimates of test positivity were rather high in Uruguay. However, after approximately two weeks, they decreased and remained low in the first 60 epidemic days (Fig. 1A). TP was positively correlated with the daily number of new cases (r = 0.75, Fig. 1B). An alternative way to monitor the progression of this pandemic is to plot the cumulative number of test-positive individuals (cases) together with the cumulative number of SARS-CoV2-positive fatalities. This approach revealed two curves, which were not parallel (Fig. 1C). Therefore, fatalities could not be predicted from the case data. A similar finding was revealed when the cumulative number of cases was compared to the cumulative number of tests performed per million inhabitants (TPMI): the curves were not parallel (Fig. 1D). One data inflection revealed that, between two and three epidemic weeks (after ~ 2000 TPMI were reached), the number of TPMI increased lin­ early over time (Fig. 1D). A similar pattern was observed when the cu­ mulative number of fatalities was compared to TPMI: after TPMI reached 1000, the number of deaths flattened out (Fig. 1E). These Fig. 2. Inferential statistics on COVID-19, Uruguay, March to May, 2020. (A). Daily test positivity against epidemic days and tests per million inhabitants; (B). Daily test positivity versus cumulative tests per million inhabitants reveal an L-shaped curve; (C). Temporal evaluation of daily test positivity reveals three distinct stage over the period, March to May 2020; (D). Cumulative cases and cumulative active cases plotted against a temporal timeline. There was a negative correlation between the daily test positivity and tests/million inhabitants (r= - 0.66 [CI= - 0.79, - 0.48]), Fig. 2A. F.O. Fasina et al.
  • 4. Methods 195 (2021) 15–22 18 patterns were also noticed when TPMI was compared to the cumulative number of deaths reported per million inhabitants (DPMI): only after TPMI exceeded 1000, the DPMI values flattened out (Fig. 1F). Several of these variables showed at least two phases (Fig. 1 C, E, F). The earliest stage displayed a quasi-exponential growth (i.e., the number of cases grew faster than time, which resulted in a quasi-vertical pattern). Later (after a data inflection was revealed) a quasi-linear phase was observed, in which the growth in the number of cases was slower than time and, therefore, cases exhibited a quasi-horizontal pattern (e.g., Fig. 1C). Test positivity was negatively correlated with testing in Uruguay (r = − 0.66, Fig. 2A). Three subsets of temporal observations showed decreasing values of TP, which corresponded to increasing (and statis­ tically significantly different) values of TPMI (p < 0.01, Fig. 2B, C). When the sub-group of active cases was compared to all cases (which included deaths and recovered cases), the cumulative number of active cases exhibited a data inflection followed by a decreasing trend which, together, supported the view that patterns displayed by the data on active cases may inform earlier than the alternative indicator (Fig. 2D). Test positivity was positively correlated with deaths/million in­ habitants (DPMI) when the six countries were simultaneously investi­ gated (r = 0.65, Fig. 3A). However, when the same countries were investigated in terms of TPMI and TP, a distinct, non-linear pattern was detected (Fig. 3B). When one seemingly outlier country was removed from the analysis, a negative correlation was revealed (r = − 0.74, Fig. 3C). When DPMI and TPMI were investigated, a non-linear (L-sha­ ped) distribution was noticed (Fig. 3D). When time, TP, and DPMI were plotted, three patterns were detected, which were characterized by a) low TP and low DPMI (a pattern displayed by three countries), b) in­ termediate values (shown by two countries), and c) high TP and high DPMI, which were only revealed by one country (Fig. 3E). In contrast, when TPMI was plotted together with time and TP, four patterns were Fig. 3. External validity and comparison of observed patterns for COVID-19 in six countries. (A). Deaths per million inhabitants against test positivity; (B). Tests per million inhabitants against test positivity; (C). Tests per million inhabitants against test positivity, excluding Chile; (D). Deaths per million inhabitants against tests per million inhabitants, excluding Chile; (E). Three-dimensional plot of deaths per million inhabitants against temporal scale and test positivity; (F). Three- dimensional plot of tests per million inhabitants against temporal scale and test positivity. There was a positive correlation for five countries except Chile, between the test positivity and deaths per million inhabitants (r= 0.65 [CI= 0.44, 0.80]), Fig. 3A. However, there was a negative correlation between test positivity and tests per million inhabitants after excluding Chile (r= - 0.74 [CI= - 0.86, - 0.55]). This demonstrates that TP may be a reproducible metric, provided that the context and other variables are also considered. F.O. Fasina et al.
  • 5. Methods 195 (2021) 15–22 19 observed, which revealed a) high TPMI and low TP values (found in two countries), b) a lower TPMI but a higher TP than the previous subset (a profile shown in one country), c) the lowest TPMI and highest TP per­ centages (detected in two countries), and d) high TPMI and high TP values (a pattern observed in one country, Fig. 3F). When geography was considered, the association between TPMI and DPMI was further documented (Fig. 4A-F). Within two weeks, one country partially predicted the fatalities associated with epidemic dispersal: the lowest TPMI values preceded an ~250% growth in DPMI. However, exceptions to this pattern were also noticed: in the same period of time, two countries that also displayed low TPMI values showed a negligible change in DPMI (Fig. 4A-F). The temporal fluctuation of TP values was minimal across countries. In the period under study and without exceptions, countries showed either one- or two-digit percentages (Fig. 5A-F). In addition to TP-related geo-temporal assessments, the shape of the data on active cases was also informative. For instance, the detection of data inflections could be viewed as a positive outcome (Fig. 6A–C). Similarly, both the directionality and numerical values of the ratio be­ tween recovered and active cases could be considered: when the ratio is greater than 1, a positive prognosis may be considered. In addition, when time progresses faster than the epidemiologic metric (when a horizontal directionality predominates) a rather stagnant situation may be inferred. In contrast, when the ratio of recovered individuals over all active cases grows faster than time (when a vertical directionality is noticed), a positive prognosis is defensible (Fig. 7A-C). 4. Discussion Hoping to explore a novel metric –test positivity− , we evaluated Fig. 4. Country-level comparison of cases, mortality and test positivity per million inhabitants for COVID-19. Plot of deaths per million inhabitants against tests per million inhabitants in (A). April 24, 2020; (B). April 27, 2020; (C). April 28, 2020; (D). May 4, 2020; (E). May 6, 2020; (F). May 9, 2020. Fig. 4 (A – F) show a synopsis of a geo-temporal analysis; it includes (low-resolution or aggregate) geographical data and a temporal description of epidemics that started less than 6 weeks apart. For visual comparisons, each plot is divided into 4 quadrants (’low & low’, ’high & low’, ’low & high’, and ’high & high’) It is shown that high TP is associated with a faster growth of deaths/mill inh. The opposite pattern (low test positivity & low deaths/mil inh) shows one exception. F.O. Fasina et al.
  • 6. Methods 195 (2021) 15–22 20 several epidemic parameters associated with SARS CoV-2 for the first 60 to 90 epidemic days in six countries where the time when the first COVID-19 case was reported differed in six or fewer weeks. Given that test positivity is not a direct measure of health status at the population level, we wondered whether this metric could estimate –albeit indi­ rectly–the progression (or lack) of epidemic dispersal. The answer to such a question is unlikely to be totally elucidated with a single study. However, one study may provide information that supports or rejects theories and/or technologies aiming at monitoring population health status when exposed to infectious diseases with a high or very high percentage of asymptomatic individuals. Earlier studies have shown that test positivity predicts epidemic dispersal better than the simple number of tests performed per million inhabitants [5,6]. In addition, this evaluation documented several sta­ tistically significant associations that may reflect biological relation­ ships. For instance, when two sources of data were used, the Uruguayan case revealed a negative correlation between TP and TPMI, both when it was investigated alone and when four other countries were explored (Figs. 2A and 3C). Such a reproducibility (which involves two data sources and five countries) does not support random findings. The fact that several variables also exhibited reproducible patterns (such as two temporal phases), provides a group of metrics associated with TP that may be worth considering in future studies. Some of these indicators may provide geography- (country-), and time-specific infor­ mation potentially useful in epidemic monitoring. For instance, in the Uruguayan epidemic, at least 1000 tests per million inhabitants appeared to be needed to flatten out the growth of fatalities (Fig. 1E). In all six investigated countries, the simultaneous consideration of TPMI and DPMI, over time, may help to find both similarities and differences (Fig. 3E and F). The observed relationships among TP, tests/million inhabitants, the shape of active cases, and the shape of the recovered/active cases (R/A) ratio support the WHO’s recommendations on policies aimed at low TP percentages: the highest levels of testing (which may detect credible levels of disease prevalence) only occurred at the lowest levels of TP. The test positivity, together with the shape of the number of active cases, can determine whether and when the epidemic is/is not under control. Associated patterns –such as increasing or decreasing trends of the recovered/active ratio data–may inform policy makers whether an overflow of hospital beds and associated resources is/is not likely in the following weeks. When TP is analyzed together with tests/million in­ habitants, it may indicate (in a country-specific manner and without pre- Fig. 5. Selected country classification by cases, mortality and test positivity for COVID-19. Plot for (A). Uruguay; (B). Chile; (C). Cuba; (D). Bolivia; (E). Argentina; (F). Mexico. Fig. 5 (A – F) show the same data depicted in fig 4, now at individual countries. The left column (A, C, E) displays countries with one digit of test positivity percentages. The right column (B, D, F) shows two-digit test positivity percentages. F.O. Fasina et al.
  • 7. Methods 195 (2021) 15–22 21 established assumptions) whether testing is sufficient or not. While the data analyzed in this study referred to nationwide levels and aggregated data tend to be less informative because they may lose resolution [17], these metrics could also be used with lower-scale, higher-resolution (county- or city-level) geographical data [11,12]. While limited in scope, this study generated hypotheses. One example is the hypothesis that lack of explicit isolation policies may inhibit the effectiveness of massive testing. Such a hypothesis was illustrated by one country that showed a high level of testing, but did not pursue isolation [18]. Patterns observed were compatible with studies that have reported (a) lockdowns but no specific testing scheme, and (b) neither testing nor lockdowns [18]. Thus, this initial study on test positivity provides support for several recommendations. We advocate for a policy that focuses on four aspects: (1) massive and repeated testing, (2) border control, (3) tracing of every case, and (4) active isolation which, to be successful, must be imple­ mented within a Critical Response Time, that is, before the exponential growth kicks in [19]. Effective responses are not likely to depend on piecemeal approaches that only consider one or a few strategies. Instead, new and accelerated educational programs are needed, which should provide training on several disciplines, including but not limited to (a) geography, (b) epidemiology, (c) the economics of public health, (d) development and use of new software packages that facilitate early (real time) and geographically specific interventions, and (e) the creation of new systems that both collect and disseminate data. Provided that additional studies reproduce these findings, one possible application of test positivity is to become an alternative to the Fig. 6. Temporal-related metrics in the analysis of COVID-19 cases in three countries. (A) Argentina; (B) Cuba; (C). Uruguay. Fig. 7. Plots of temporal pattern of the ratio between recovered and active cases in selected countries. (A) Argentina; (B) Cuba; (C). Uruguay. F.O. Fasina et al.
  • 8. Methods 195 (2021) 15–22 22 quasi-centennial ‘reproductive number’ (or R0), which estimates the ratio of secondary over primary cases. That is so because, at the present time, there are no real time estimates on how rapidly epidemics may progress and such estimates are also prone to numerous sources of error [20,21]. Unlike the lack of geographically explicit information and delayed calculations associated with the R0 (which prevent real time, site-specific decision-making), test positivity can be calculated on daily basis and provide actual (not assumed) high-resolution, geographical information (e.g., neighborhood- or village-specific geographical co­ ordinates). Such an application could improve policy-making: instead of assuming test positivity is homogeneously distributed over large terri­ tories and remains constant over time, daily changes at specific (and geographically small) sites could support context-specific, cost-benefit oriented interventions [6]. To win the war against the next epidemic, reactive responses are not acceptable. Proactive responses are the only alternative, which require a robust and immediate creation of novel educational programs of inter/ transdisciplinary nature. To that end, further research on the advantages and limitations of test positivity is necessary. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Appendix A. Supplementary data Supplementary data to this article can be found online at https://doi. org/10.1016/j.ymeth.2021.05.017. References [1] D.P. Oran, E.J. Topol, Prevalence of asymptomatic SARS-CoV-2 infection A narrative review, Ann Intern Med 173 (2020) 362–367, https://doi.org/10.7326/ M20-3012. [2] A.J. Ing, C. Cocks, J.P. 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