diagnostic micro and lab methods

Question 1

bright field

Answer 1

1) light illuminates

Question 2

dark field

Answer 2

1) spirochetes can be seen

Question 3

fluorescence

Answer 3

1) fluorescent labels or bacteria itself is fluorescent

Question 4

gram stain

Answer 4

1) gram positive does not pick up safranin because it should be stained purple from the crystal violet in peptidoglycan

Question 5

acid fast stain

Answer 5

1) stained red by carbol fuchsin, retained after decolorization
- methylene blue stains remaining organinisns
2) mycobacteirum

Question 6

immunoflucoresence

Answer 6

1) antibody that binds to the HSV1/2
2) enzyme immunoassay for antigen localization in cells
3) direct
- primary antibody probe
4) indirect
- primary antibody and secondary antibody is tagged

Question 7

culture media should include

Answer 7

1) components for the growth of certain types of bacteria
2) ex. blood agar for beta hemolytic bacteria (yellow) and alpha hemolytic bacteria
- H2O2 to oxidizes the Fe2+ in HgB to F23+ metahemoglobin (green)

Question 8

identify fungi with buffered charcoal, yeast extract

Answer 8

1) legionella
2) sabouraud dextrose agar is useful in isolation and growth of fungi like candida

Question 9

selective media

Answer 9

1) recover particular microorganisms from a mixed organism
2) staph?

Question 10

analytical profile index

Answer 10

1) 20 mini test chambers having chemically defined compositions for each test

Question 11

restriction endonucleases

Answer 11

1) ecoRI
- natural products of bacteria
- protect from viral DNA
- cut it at specific sequences
- RE that cuts outside GOI and cut vector plasmid with same RE
2) then the gene can be cloned

Question 12

cloning vectors

Answer 12

1) plasmid has ampicillin resistance gene
2) site has a polylinker

Question 13

transformation of bacteria

Answer 13

1) only cells with the correct plasmid will survive on ampicillin

Question 14

screening

Answer 14

1) phenotypic screening
- different color or morphology
2) screening with antibodies

Question 15

cloning eukaryotic genes

Answer 15

1) eukaryotic DNA has noncoding regions
- introns
2) after mRNA is transcribed the introns are removed
3) reverse transcribe
- make DNA copy of the mRNA
- cDNA

Question 16

cDNA cloning

Answer 16

1) mRNA isolated from the cell
- RT and dNTPs
2) cDNA amplified and inserted into a vector

Question 17

frederick sanger

Answer 17

1) sequencing organisms based on DNA sequence
2) use of ddNTPs
2) electrophoresis of varying length fragments
4) automated
- fluorescent markers
- lase causes fluorescence and detector determines nucleotide

Question 18

capillary electrophoresis

Answer 18

1) can sequence DNA that way
2) using electric field

Question 19

PCR

Answer 19

1) primers
2) taq polymerase
3) heating and cooling
4) dNTP

Question 20

real time PCR

Answer 20

1) fluorescence that shows the amplification of DNA

Question 21

ELISA

Answer 21

1) immunoassays for quantitation of antibody or antigen
2) antibody detection
- antigen coated wells
3) antigen capture
- antibody coated wells and detection by secondary antibody

Question 22

bacteria hybridization?

Answer 22

1) culture grown
2) extract DNA and do stuff to make cDNA
3) label with fluorescence
4) mix and hybridize to genes