C6 Flashcards

1
Q

Application of molecular detection

A
  • Bacteria
  • Virus
  • Fungi
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2
Q

Explain respiratory tract pathogens

A
  • Easily transmitted by contact with infected person
  • Lab detection lack sensitivity & time consuming
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3
Q

Explain M.tuberculosis

A
  • Diagnosis took long time (spread the disease)
  • Diagnose by mycobacteria smear & culture
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4
Q

Example of MTB methods

A
  • Amplified M.tuberculosis Direct Test
  • AMPLICOR M.tuberculosis PCR Test
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5
Q

Explain amplified M.tuberculosis direct test

A
  • 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
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6
Q

Explain AMPLICOR MTB Test

A
  • 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
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7
Q

Time and sensitivity of Amplified & AMPLICOR

A

Ampified
- 3.5 hours
- Positive: 100%
- Negative: 83%

AMPLICOR
- 6.5-8 hours
- Positive: 94.7%
- Negative: 55.3%

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8
Q

Explain detection of virus

A
  • Detect by testing antibodies, measure presence or absence of viral antigen, detect growth
  • Viral antigen detect by enzyme immunoassay or direct immunofluorescent assay
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9
Q

Explain molecular diagnostic in virus

A
  • Use PCR, RTPCR, qPCR, TMA & bDNA to diagnose viral infections
  • HIV, HCV & HPV use molecular based test
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10
Q

Example of nucleic acid test for virus

A
  • HIV: PCR, RT-PCR, bDNA
  • HPV: PCR, qPCR
  • Corona & Influenza: RT-PCR
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11
Q

Test performance features for viral load measurements

A
  • Sensitivity: lowest level detect at 95% of the time
  • Accuracy: closeness measure value to standard
  • Specificity: negative & positive result true
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12
Q

Detection of resistance & virulence

A
  • 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
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13
Q

Mechanism of resistance

A
  • Destruction of agent: Beta lactamase
  • Elimination of agent: Multidrug efflux system
  • Alter cell wall structure
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14
Q

4 reason for using molecular based method to detect resistance

A
  • 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
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15
Q

Explain molecular epidemiology

A
  • 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
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16
Q

Epidemiology typing methods

A

Phenotypic
- Serotyping
- Biotyping

Genotypic
- Ribotyping
- Plasmid analysis