Dr. Stillwell Molecular Biology Notes

Question 1

virology is viral detection using what two methods?

Answer 1

Culture or PCR

Question 2

How can we interpret real-time polymerase chain reaction (qPCR)? ** ON EXAMMM

Answer 2

** Positive/negative with a CT value (lower CT value means more DNA copies present in the sample)

Question 3

***explain fluoresce vs cycle number on the graph representing one cycle of qPCR:

Answer 3

Different number lines are different samples run
Each sample must pass “threshold” to be considered positive (point at which it passes threshold is called the CT value) *Cycle number at which sample crosses the diagnostic threshold KNOW FOR THE EXAMMMM

Question 4

*** ON EXAM What does Histology detect?

Answer 4

***Pathogen morph or viral inclusions

Question 5

*** ON EXAMMM what does PCR and qPCR detect??

Answer 5

Nucleic acid (viable or not)

Question 6

*** ON EXAMMM What does culture detect?

Answer 6

Live/viable pathogens ONLY

Question 7

**ON EXAMMM What does Electron microscopy detect?

Answer 7

Pathogen ultrastructure

Question 8

IHC uses ___ whereas ISH uses _____

Answer 8

IHC detects proteins whereas ISH detects nucleic acids (DNA)

Question 9

IHC=

Answer 9

proteins sought out

Question 10

Advantages and disadvantages of PCR

Answer 10

Advantages- highly sens, use all kinds of prepped tissues
Disadvant- only detects presence of nucleic acid, cannot ID pathogen viability and detects only known targets, and highlyyy senst to contamination

Question 11

How is real time PCR (Aka Quantitative PCR aka QPCR different from conventional PCR?

Answer 11

Uses primers plus DNA- based dyes or fluorescent probe(s)= increased specificity
reveals QUANITY of target sequence in a sample
Results can interpret extent of infection and clinical relevance

Question 12

Lower CT value=

Answer 12

more DNA copies present in the sample

Question 13

Pathogen morph and suggestion of viral inclusions=

Answer 13

Histology