quality control: molecular diagnostics
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Quality Control: Molecular Diagnostics. Overview. Sample Collection, Transport, and Processing maintain Nucleic acid integrity through these processes Contamination Control - PowerPoint PPT PresentationTRANSCRIPT
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Quality Control: Molecular Diagnostics
Molecular Diagnostics Quality Control - Module 8 22
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)
Molecular Diagnostics Quality Control - Module 8 33
Quality Control: Molecular Diagnostics
Molecular Diagnostics Quality Control - Module 8 55
Real Time PCRSAMPLE
PRIMER
Taq polymerase Amplified Target NA
SYBR GreenQuantitative Fluorescence
+ - In
Molecular Diagnostics Quality Control - Module 8 66
Amplification Reactions
Amplification / Time
POSITIVE
NO AMPLIFICATION
Molecular Diagnostics Quality Control - Module 8 77
Real Time RT-PCR
Molecular Diagnostics Quality Control - Module 8 88
A/H3
Molecular Diagnostics Quality Control - Module 8 99
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
testpositive controls are “in-house”negative controls are “in-house”
interpretation of tests need to be done with caution