ORAL PRESENTATION MADE BY ASARE, KUMI KWAME AT: The Sixth Ghana Biomedical Convention Biomed – 2013 conference held at University of Cape Coast; Cape Coast, Ghana from July 29TH – 31ST 2013

1. Evidence of continuous use of Chloroquine
(CQ) in Ghana after seven years of its ban, its
consequences on CQ resistant markers
ASARE, KUMI KWAME
Department of Biomedical and Forensic sciences,
University of Cape Coast
“you read about it in the news but you don't believe it you'll only know
about it when the man in the long black coat knocks on your door 'cause
you're his next victim”…Lucky Dube

2. Introduction
K. K. Asare, et al., 2

3. CQ resistance
Late 1950s and early 1960s
In 1965 CQ resistance had been alleged by
Schwendler (Beausoleil, 1967).
CQ resistant P. falciparum were isolated in
1986 (Neequaye, 1986)
K. K. Asare, et al., 3

4. In 2001, CQ resistance P. falciparum in Ghana was
reported to be around 88% (Evans et al., 2005)
In 2005,MoH finally implemented ACTs
Despite the replacement anecdotal evidence
suggests that CQ is still in used in many communities
in the country.
K. K. Asare, et al., 4

5. The continuous use of the former drug would result
in development of stable mutations in:
I. Pfcrt gene
II. Pfmdr-1 gene
Which can lead to cross-resistance to other
antimalarial drugs such as:
1. Amodiaquine
2. Artemisinin
3. Quinine
K. K. Asare, et al., 5

6. Aim & Objectives
K. K. Asare, et al., 6

7. Methodology
Solomon-Saker method
I.0.05%TBPEE-in-chloroform
Molecular method
I.Chelex dna extraction
II.Nested PCR methods
III.RFLP
Statistical analysis: SPSS-
Chi-square at 95% CI
K. K. Asare, et al.,
STUDY SITES
SUBJECT SELECTION
MALARIA INFECTED
PATIENTS
NON-MALARIA
INFECTED PATIENTS
SUBJECTS WHO
WERE NOT
ENROLLED
ENROLLED SUBJECTS
SAMPLE TAKEN
BLOOD SAMPLE URINE SAMPLE
MYSTERY BUYING
METHOD
SALES OF CQ
BY COMMUNITY
PHARMACY &
CHEMICAL SHOPS
1. HAEMATOLOGICAL ANALYSIS
2. MOLECULAR ANALYSIS
1. URINALYSIS
2. URINE CQ ANALYSIS
1. ELMINA HEALTH CENTRE
2. CAPE COAST DISTRICT HOSPITAL
3. TWIFO PRASO DISTRICT HOSPITAL
4. ST. FRANCIS XAVIER HOSPITAL, ASSIN
FOSO
ETHICAL CLEARANCE:
1. GHSERC
2. UCCIRB
QUESTIONNAIRE
ADMINISTRATION
1
2
7

8. RESULTS AND DISCUSSION
K. K. Asare, et al., 8

9. K. K. Asare, et al.,
 Fig.1: Samples of chloroquine injections
1. They were mainly manufactured at India & China
2. About 20% of CQ injections obtained did not have labels
FIG. 1
9

10. K. K. Asare, et al.,
 Fig.2: controls samples fortified with CQ and patients samples that contained CQ
 Cross-reactivity test were performed
 Contrary to Malawi
 Asia & South-America case, Ghana has maintained high CQR due to continuous
CQ usage
FIG 2
10

11. K. K. Asare, et al.,
 Fig. 3: Apo 1 RFLP at K76T in pfcrt
 Persistent CQ use subject the parasite to drug pressure
 Development of stable T-76 mutation & other set of mutations in pfcrt
gene
200
100
234
FIG. 3 FIG. 4
11

12. K. K. Asare, et al.,
 Fig. 5: Dpn II RFLP at D1246Y in pfmdr-1
 Mutations in pfmdr-1 is synergetic to pfcrt in CQR mechanisms
 Combination of haplotype mutations in pfmdr-1 modulates CQR
800
400
100
FIG. 5 FIG.6
12

13. • 86 Y & 184 F
mutations play a role
in CQR
• Studies from South-
America, Asia &
Africa have shown
that 86 Y & 184 F
confer resistance to
QN, MFQ &
Halofantrine
• Also modulate
parasite sensitivity to
artemisinin drug
K. K. Asare, et al., 13

14.  Initial studies
reported
association
between 86 Y in
pfmdr-1 & CQR
 However, several
field studies
showed
inconsistency
 Again,
transfection
studies showed
that replacement
of 86 Y with 86 N
in pfmdr-1
decrease CQR
from high to
moderate levels K. K. Asare, et al., 14

15. CONCLUSION & RECOMMENDATION
 The study has revealed continuous use of CQ in the country
 Increase of point mutations in pfmdr-1 gene
 These mutations can cause cross-resistance to current anti-
malarial drugs
We recommend:
a. Stringent regulations
b. Further studies:
1. To investigate CQ importation
2. Drug sensitivity and pharmacokinetics studies to validate our
findings
K. K. Asare, et al., 15

16. K. K. Asare, et al., 16
Acknowledgements
 DR. JOHNSON N. BOAMPONG
 DR. NEILS B. QUASHIE
SUPERVISORS
SCHOOL OF BIOLOGICAL SCIENCES
Department of Biomedical and Forensic sciences