The document discusses the history and increasing importance of automation in microbiology laboratories, particularly for blood cultures, noting that traditional culture methods can take 72 hours for results while rapid automated methods are needed to identify pathogens faster to guide antibiotic treatment for infections like sepsis. It also outlines the steps involved in optimal blood collection for cultures, including patient preparation, materials, and techniques to maximize success in collecting pediatric samples.

1. AUTOMATION IN
MICROBIOLOGY
(for blood cultures)
Dr.T.V.Rao MD

2. Beginning of Microbiology

Almost exactly 300
years ago Anton van
Leeuwenhoek
described the first
bacteria seen through
the microscope, thus
providing the technical
basis for studying the
morphology of microorganisms

3. Knowledge explosion in
Microbiology

The explosion of knowledge in the last
century, pioneered by Pasteur, Koch and
many others immortalised in modern
generic and specific names was
dependent on improvements in
procedures for isolating and identifying
organisms of importance in industry and
medicine.

4. Rapid Methods are
Emerging

Rapid methods and automation is a
dynamic area in applied microbiology
dealing with the study of improved
methods in the isolation, early detection,
characterization, and enumeration of
microorganisms and their products in
clinical, food, industrial, and
environmental samples.

5. Changing perceptions ,

Microbiology labs are beginning to accept
the shift toward automation for reasons as
numerous and varied as microbiology
itself. Microbiologists and lab technicians
recognize that automated solutions are
not intended to replace cognitive decisionmaking but rather, simply replace tedious,
repetitive steps.

6. Man has Evolved So also the
Microbes, so the need for
Automation

7. Beginning of Automation


The field started around mid-1960s and
with the development of a variety of
miniaturized microbiological techniques
developed
Streamlining workflow maintains
consistency but allows microbiologists to
devote more time to operations that require
their unique skills and experience

8. Hospital Acquired Infection a threat to
Medical Profession- needs faster
methods to Identify.

Microbiology labs play a crucial role by
establishing a front line of defence against
the threat of Nosocomial infections.
According to the Centres for Disease
Control and Prevention, in American
hospitals alone, HAIs account for an
estimated 1.7 million infections and
99,000 associated deaths annually. Of
these infections, 32% are urinary-tract,
22% are surgical-site, 15% are
pneumoniae, and 14% are bloodstream.

9. The Clinicians need Quicker
Results

The shift from
manual test
processing to
automated solutions
can aid in reducing
HAIs by providing
health systems with
standard, consistent
lab processing that
yields quicker, more
accurate test results.

10. Automation enters into several
areas in Microbiology

Rapid Methods and Automation in
Microbiology has developed into an
important sub-discipline of applied
microbiology in the past 15 years. The
field deals with improved methods in the
isolation, early detection, characterization,
and enumeration of microorganisms and
their products in clinical, food, industrial,
and environmental samples.

11. Automation reduces errors and
innovative

Automated processes have gone well
beyond changing outdated procedures.
They minimise potentially dangerous
practices, lower turnaround time, reduce
errors, enhance quality control, improve
specimen handling and boost accuracy.
Moreover, technologists in automated
labs tend to embrace innovation more
readily.

12. Lesser Manpower More volume
of work

Because automation largely circumvents
repetitive manual processing, lab
technicians find it easier to focus on
complex tasks that require their specific
skills.
It is therefore remarkable that, despite
shortages of skilled personnel and
increases in the volume of work, it has
taken years for automation to become
acceptable to microbiologists.

13. Desired Objectives in Automation

Reexamination of laboratory functional steps
–
–
–
–


Phlebotomy
Sample labeling
Transportation
Pre-, peri-, and post-analytical processing
Laboratory automation for improved efficiency and
error reduction
Create an informatics continuum
– Process Control vs. LIS function
– Auto-verified and auto-interpreted data

Predictive genomics and the passive home
monitoring paradigm

14. Adding Value to Lab Tests Through Automation

Lab Test
– Faster TOT
– Accuracy,
Precision, Safety

Add information
value
– Auto validation
– Trending

Effecting
change using
lab results
– Lifestyle
changes
– Selection of
therapeutics
Lab Test
Auto validation
Trending
Life Style Adjustments
Appropriate Therapeutics

15. Every Body is a Learner to New
Technologies, Ent hus i as m
Makes Difference

16. Blood culturing most
important and live saving
Investigation
Needs optimal Methods for
Diagnosis of Blood Borne
Pathogens

17. Why Blood Culture
 Physician
infection
can find source of
 Physician
can initiate lifesaving support measures
 Physician
can start targeted
antibiotic therapy

18. What is a Blood Culture?

A blood culture is a
laboratory test in
which blood is
injected into bottles
with culture media to
determine whether
microorganisms have
invaded the patient’s
bloodstream.

19. Need for Blood Culture?
No microbiological test is more essential to the
clinician than the blood culture. The finding of
pathogenic microorganisms in a patient’s
bloodstream is of great importance in terms of
diagnosis, prognosis, and therapy.”
- L. Barth Reller, Clin. Infect. Diseases, 1996

20. A
Proof in Blood borne
Infection
clinically suspected infection is
ultimately confirmed by isolation or
detection of the infectious agent.
Subsequent identification of the
microorganism and antibiotic susceptibility
tests further guide effective antimicrobial
therapy. Bloodstream infection is the most
severe form of infection and is frequently
life-threatening, and blood culture to
detect circulating microorganisms has
been the diagnostic standard.

21. Blood culture is a Important a
Diagnostic tool in Infectious
diseases
 Blood
culture is a microbiological
culture of blood. It is employed to
detect infections that are spreading
through the bloodstream
(such as bacteraemia, septicaemia
This is possible
because the bloodstream is usually
a sterile environment.
amongst others).

22. What are We doing Now is not
optimal – Needs Rapid Methods
 Most microbiological culture procedures
require the use of solid media, like blood
agar and MacConkey agar plates that
need to be visually monitored by trained
personnel at intervals of 24 hours. These
conventional cultures using normal media
take at least a minimum of 72 hours to
isolate the pathogen and carry out
susceptibility test to know the efficacy of
antibiotics on simple aerobic bacteria

23. Optimal Methods of Blood
Collection makes difference





The physicians consent with filled in request with details
for culturing the Blood
Gloves will be worn in accordance with standard
precautions..
Appropriate verification of the patient's identity, by
means of an armband or area specific procedure, will
occur before the specimen collection.
Cultures should be drawn before administration of
antibiotics, if possible.
If at all possible, blood cultures should not be drawn from
lines, but should be drawn only via venepuncture

24. PHLEBOTOMY TRAINING
PROGRAMS
WHO?
WHAT?
WHERE?

25. NEW CATEGORY OF
LAB PERSONNEL
Phlebotomist
Defined as a person who collects blood for
clinical laboratory test or examination
purposes


26. Principles for Collection





Gloves will be worn in accordance with standard
precautions.
•A physician’s order must be obtained for specimen
collection.
•Appropriate verification of the patient's identity, by
means of an armband or area specific procedure, will
occur before the specimen collection.
•Cultures should be drawn before administration of
antibiotics, if possible.
•If at all possible, blood cultures should note drawn from
lines, but should be drawn viavenipuncture.

27. Materials











Chlorhexidine swabs (1-2 packages)
Alcohol swabs
Blood culture bottles (2 bottles per set)
2 syringes (adult: 20 cc, paediatric: 5 cc)
2 needles (adult: 22 gauge or preferably larger butterfly
or standard needle; pediatric: 25 or 23 gauge butterfly or
standard needle)
Gloves (sterile &nonsterile)
Tourniquet
Sterile gauze pad
Adhesive strip or tape
Self-sticking patient labels
Plastic zip lock specimen bags

28. Steps 1 – 3, Check, Explain,
Wash



1.Identify the patient by
checking the arm band or
area-specific procedure.
2.Explain the procedure
to the patient.
3.Wash hands with soap
and water with friction for
15 seconds or use
alcohol based hand rub

29. Step 4 –Prep Cap

Prep the rubber cap
of the blood culture
bottles with an alcohol
pad in a circular
motion. Allow the
alcohol to dry.

30. Step 5 -Prep the Puncture Site

Prep the puncture site with Chlorhexidine:
– •Using aseptic technique, remove the applicator from
its package.
– •Holding the applicator downward, gently squeeze the
wings to release the solution.
– •Scrub with a back & forth motion using friction for 30
seconds on dry skin or 2 minutes on wet skin.

•Do not wipe the site after cleansing the skin
with Chlorhexidine.

31. Step 6 -Gloves






Apply gloves:
If palpation of site prior
to puncture is
anticipated, wear sterile
gloves.
If palpation of site prior to
puncture is not
anticipated, wear
nonsterile gloves.

32. Step 8 -Mix

Gently rotate the
bottles to mix the
blood & the broth (do
not shake vigorously).

33. Step 9 and 10 (Label)


Place the patient label on each bottle & label
each culture bottle with the site of specimen
collection. When applying patient identification
labels, do not cover the bar code label on the
blood culture bottles. Attach the laboratory
requisition.
Send the blood cultures to the Clinical
Microbiology receiving area as soon as possible.

34. Step 11
 11.Document
the following in the medical
record
Date & time specimen obtained
 –Site of specimen collection
 If 2 sets of blood cultures have been
ordered, obtain the second set in the
same manner as the first, making a new
venepuncture at a different site

35. Techniques of Paediatric Collections
Tourniquet Application





Need to maximize chances of successful
collection
Remember that the vein is still developing and
might need to rely on firm tightness
Ideally, tourniquet should not be kept on for
more than one minute
If possible, apply heat
If using a hand, consider a bucket of warm
water

36. Techniques of Paediatric Collections
Insertion Principles


Cantilevering of elbow
The option factor:
-Choose your options of direction before insertion
-Minimize the odds of unnecessary “digging”



Avoid plunging the needle right up to the hilt
Often, a drawback does the trick
If vacutainer is slowing down, replace with a
syringe

37. Self Protection
A few ways to make sure your
role in the collection process is
carried out with efficiency,
orderliness and safety

38. The Contaminated Blood
Culture


If the skin is not adequately cleansed before
drawing blood for culture, bacteria on the skin will
be injected into the bottle, producing a false
positive blood culture.
It is sometimes difficult for the physician to
determine whether the bacteria growing in the
blood culture is a real pathogen causing
bloodstream infection or whether bacteria on the
skin have contaminated the culture. This can lead
to excess use of antibiotics and prolongation of
hospital stay.

39. Sample Labeling
Efficiencies
 Bar coding at the point-
of-phlebotomy
 2D vs. 1D bar codes
– Reduce the number
of computer
interfaces
– Self directing
specimens
B-D id

40. Technological Improvements for All
Steps in the Diagnostic Process
Just in Time Supplies
Automated
Phlebotomy
Trays
Process Control
EMR
Reporting
Instruments
Designed for
Automation
Storage
RFID
2D-Codes
Recording
Biorepository
Transportation
Analysis
Pre-analytical
Analytical and
Automation
Mobile Robot
Accessioning

41. What is a Blood Culture?

A blood culture is a
laboratory test in
which blood is
injected into bottles
with culture media to
determine whether
microorganisms have
invaded the patient’s
bloodstream.

42. Blood & Body Fluid Cultures




Blood cultured by the BacT/Alert 3D leads to early
detection of pathogens (>89 per cent within 24 hours
and 97 per cent within 48 hours) especially in cases of
septicaemia, enteric fevers, bacterial endocarditis and
other pyrexias of bacterial origin.
Activated charcoal neutralises antimicrobials and toxins
enhancing early recovery of pathogens. Positives are
detected faster than Bactec even at low concentrations
in blood and body fluids like CSF, CT guided aspirates
etc.
Delayed transport does not compromise results.
The instrument is capable of recovering significantly
more organisms that resin.

43. BacT/AlerT 3D culture
system
 BacT/AlerT 3D culture system. This is the
first automated non-radiometric and noninvasive culture system that continuously
monitors system for culture of bacteria
(both aerobic and anaerobic), fungi and
mycobacteria. All these bacteria can be
cultured using different media as
prescribed..

44. Principles in BacT/AlerT 3D
culture system
 This is a closed system and works on the
colorimetric principle of detection of CO2
produced by the organisms. The CO2
causes a lowering of the pH of the
medium, which in turn produces a colour
change in a sensor attached to the CO2sensitive base of each bottle.

45. You are guided by
Computerized Systems
 The instrument reacts before this colour
change is apparent by means of an
audible or visible alert flagged by the
computer. The bottles are constantly
agitated and are read at 10-minute
intervals. The readings are transmitted to
a computer compiler, which computes
results. This

46. bioMérieux BacT/ALERT® 3D
 The bioMérieux BacT/ALERT® 3D provides
an optimal environment for the recovery of a
wide range of pathological organisms,
including bacteria, yeasts and
mycobacteria; utilizing proprietary plastic
culture bottles ensuring added safety to the
user.

47. Principles of functioning of BacT alert
Monitors



Microorganisms multiply
in the media, generating
CO2. As CO2 increases,
the sensor in the bottle
turns a lighter colour.
Measuring reflected light,
the BacT/ALERT 3D
monitors and detects
color changes in the
sensor.
Algorithms analyze the data to
determine positivity, and the
laboratory is notified
immediately with visual and
audible alarms.

48. Automation becomes more
complex

49. Automation becomes need of the
Hour
 Full microbiology laboratory automation
needs have never been so apparent, with
financial constraints and increasing testing
volumes at the same time that labour is
becoming both harder to find and more
expensive. Implementation of full
microbiology lab automation is one
solution, as fewer technologists are
required to process automated tests..

50. Automation improves quality
of services

Overall, laboratories
transitioning from
conventional to
automated processes
find that technologists
and microbiologists
are more open to
innovation and
improved quality.

51. Industry flourishes too..

An entire industry of
microbial diagnostic kits
flourished to the present
day. Next in the 70s the
development was in
immunological test kits
and instruments to
monitor the presence of
food borne pathogens
and biomass and to
predict microbial growth
automatically.

52. Advantages of automation

Automated solutions have recently
emerged in the marketplace that address
key areas of the microbiology lab.
Automating these processes-simple,
standard, or complex-can revolutionize
the microbiology lab with more efficient,
standardized practices that will improve
quality, safety, and cost-efficiency.

53. Automation increases efficacy and
eliminates individual variations
 For example, automating small, yet vitally
important tasks, can make a huge impact
on the efficiency and accuracy of
laboratories. Lab technicians streak an
estimated ??? agar plates a day, a
process that is laborious, tedious, and
inconsistent. Each lab technician has his
own streaking technique

54. Automation is
Advantageous
 Include the elimination of subjective
variability,
 Savings in media and reagents, and the
earlier production of useful information in
many instances, all of winch can make a
substantial contribution to productivity and
the control of runaway cost escalation.

55. Automation combined with Laboratory
Information management
 The
combined use of laboratory
automation and laboratory
information management software
(LIMS) has been shown to increase
productivity, reduce human error
and improve tracking and
traceability in a microbiology lab

56. Workflow Management
Automation
Information
Data
DATA
Accessioning
 Specimen tracking
 Data logging and reporting
 Quality control documentation

PROCESS
MANAGEMENT
Sample quality
assessment
 Optimal routing and
scheduling
 Intelligent reporting


57. Bar coding, robotics and computers
 Bar coding, robotics and computers that replace
manual transcription significantly reduce data
loss and errors. Automation also makes it less
likely that plate information and patient
identification will be duplicated or transposed.
Three trends will drive laboratory automation’s
future: smaller, more-flexible analysers and
automation based on next-generation
technology, including micro fluidics, easy-to-use,
powerful software for centralised lab
management, and internet-based real-time
service for better up-time.

58. Bar coding replacing the
Manual reading – Reduces
errors
 The key to this real-time automation was
real time barcode labelling of all sample
carriers (such as bags, tubes, dishes,
bottles) provided by Kiestra's Barcode
system. This was used in conjunction with
Auto scribe's Matrix LIMS. Real time
barcode reading is known to reduce
transcription error rates to only 1 in 36
trillion characters - compared to 1 in 300
characters with manual reading.

59. Kiestra's BarcodA

Kiestra's BarcodA
automatically places
an optical barcode on
all tubes, bottles and
petri-dishes that
contains important
information such as
composition, sell-by
date etc.

60. Bar-coding helps in tracing
the errors

The barcode makes
every sample carrier
unique and recorded
meaning full
traceability for the
laboratory. Samples
are also provided with
a barcode which is
generated by Matrix
LIMS.

61. Quality replacing Quantitity
 Quality
issues are becomin g in creasin gly
important in diagnostic laboratories. The fact of
quality is no longer sufficient and we must now
develop mechanisms to assure consumers, the
public and, most importantly, ourselves of the
contin uin g quality of our service. Movin g
towards a quality -assured system is not easy,
requirin g a meticulous attention to detail in all
areas of a laboratory's workin g and
organization.

62. Automation gained the
Universal acceptance
 A further plus for lab automation is that it
promotes consistency and quality. Without
automation, lab tasks that are necessarily
repetitive can lead to inconsistent or
inappropriate ways of work and, from
there, to improper treatment, longer
patient stays, medication errors and
unwanted drug side-effects.

63. Created by Dr.T.V.Rao MD
for “e” learning Programme
Email
doctortvrao@gmail.com