C6 Flashcards
Application of molecular detection
- Bacteria
- Virus
- Fungi
Explain respiratory tract pathogens
- Easily transmitted by contact with infected person
- Lab detection lack sensitivity & time consuming
Explain M.tuberculosis
- Diagnosis took long time (spread the disease)
- Diagnose by mycobacteria smear & culture
Example of MTB methods
- Amplified M.tuberculosis Direct Test
- AMPLICOR M.tuberculosis PCR Test
Explain amplified M.tuberculosis direct test
- Use transcription mediated amplification to amplify 16S rRNA
- Amplified rRNA detect using same DNA probe in DNA probe assay & measure chemiluminescence of rRNA - DNA
- Performed in 3.5 hours in single tube with 100% specificity
Explain AMPLICOR MTB Test
- Use to detect M,tuberculosis directly
- Cell are lyse in the sample releasing mycobacteria DNA
- DNA denature & primer complementary to 584 bp region of 16S rRNA
- DNA polymerase make copy of target DNA
- Product detection using DNA probe specific
Time and sensitivity of Amplified & AMPLICOR
Ampified
- 3.5 hours
- Positive: 100%
- Negative: 83%
AMPLICOR
- 6.5-8 hours
- Positive: 94.7%
- Negative: 55.3%
Explain detection of virus
- Detect by testing antibodies, measure presence or absence of viral antigen, detect growth
- Viral antigen detect by enzyme immunoassay or direct immunofluorescent assay
Explain molecular diagnostic in virus
- Use PCR, RTPCR, qPCR, TMA & bDNA to diagnose viral infections
- HIV, HCV & HPV use molecular based test
Example of nucleic acid test for virus
- HIV: PCR, RT-PCR, bDNA
- HPV: PCR, qPCR
- Corona & Influenza: RT-PCR
Test performance features for viral load measurements
- Sensitivity: lowest level detect at 95% of the time
- Accuracy: closeness measure value to standard
- Specificity: negative & positive result true
Detection of resistance & virulence
- Microbe develop defence to antimicrobial agent
- Resistant arise from alerted target binding, active extrusion
- Single nucleotide variant in drug target or transport result in resistance
Mechanism of resistance
- Destruction of agent: Beta lactamase
- Elimination of agent: Multidrug efflux system
- Alter cell wall structure
4 reason for using molecular based method to detect resistance
- Potential effectiveness of agent
- Gene involve in resistance can be detected directly
- Monitoring spread of resistance gene
- Gold standard for validation of new phenotypic assay
Explain molecular epidemiology
- Epidemiology include collection & analysis of env, microbiological & clinical data
- Study of causative genetic & environmental factors at molecular level
- Diseases: determine genetic similarities & differences among microbiological isolate
- Lab: identify & typing infectious agents
Epidemiology typing methods
Phenotypic
- Serotyping
- Biotyping
Genotypic
- Ribotyping
- Plasmid analysis