Quality Control:

Molecular Diagnostics

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Overview
 Sample Collection, Transport, and Processing
 maintain Nucleic acid integrity through these
processes
 Contamination Control
 establish a unidirectional work flow from a DNA-
free area for reagent preparation, to a sample
processing area, to area where amplification and
detection occur
 Positive and Negative Controls
 Reference: CLSI MM3-A (Molecular Diagnostic
Methods for Infectious Diseases)

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Quality Control:
Molecular Diagnostics

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Real Time PCR
SAMPLE

Taq polymerase
Amplified
Target NA
PRIMER
Quantitative
SYBR Green Fluorescence

+ - In

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Amplification Reactions

POSITIVE

NO AMPLIFICATION

Amplification / Time

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Real Time RT-PCR

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A/H3

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A/H5

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Risks with PCR
 most RT-PCR is still done as “in-house”
or “home-brew”
 primers are not necessarily optimized to
test
 positive controls are “in-house”
 negative controls are “in-house”
 interpretation of tests need to be done
with caution

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