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Medical Microbiology > LAB FINAL > Flashcards

LAB FINAL Flashcards

1
Q

What are the diagnostic methods used for?

A

identify the pathology of a particular patient

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2
Q

What is an immunoassay?

A

analytical methods using antigens/antibodies to determine the presence of their corresponding antigen/antibody in different media

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3
Q

What are immunoassays used for?

A

find the presence of specific antibodies that are produced in the response to a non-self protein (antigen)

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4
Q

Why are immunoassays useful in the diagnostic field?

A

they are rapid, cost-effective, sensitive and specific for particular antigens

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5
Q

What are the 3 main parts of a Western Blot assay?

A

separate antigens by weight | transfer (blot) to a membrane | test for antigen via enzyme-substrate reaction

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6
Q

What is the weakness of Western Blot assays?

A

less sensitive compared to others

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7
Q

What is an ELISA?

A

enzyme linked immunosorbent assay (ELISA) 96-microtiter plate capture specific antibody/antigen in sample

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8
Q

What are the 3 main steps of ELISA?

A

absorb protein-interest into wells (solid-phase support system) | use antibody-specific to protein | develop using enzyme-substrate reaction

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9
Q

What are the 3 most common ELISAs?

A

direct, indirect, sandwich

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10
Q

What is a strength of ELISAs?

A

fast, cheap, easy to perform, more sensitive

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11
Q

What is a weakness of ELISAs?

A

false positives

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12
Q

What are ELISAs used to determine?

A

ability of an antigen to bind to extracellular matrix proteins

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13
Q

What is an agglutination assay?

A

ability of antibody to attach to specific antigen = create a lattice = clumping of particles results in visible conglomerate (aggluntination)

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14
Q

What is the agglutination assay commonly used to test for?

A

Syphilis in diagnostic laboratories (VDRL assay)

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15
Q

What is a significant step of the agglutination assay? Why?

A

dilute antigen and antibody bc agglutination can only occur when the concentration of antibody and antigen is equal = zone of equivalence

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16
Q

What happens if there is an excess of antigen in the agglutination assay?

A

agglutination will not occur = leads to false positives

17
Q

What is the precipitation assay?

A

uses insoluble complex formed between antigen/antibody

18
Q

How are results observed in a precipitation assay?

A

aggregate precipitates out of solution = visualization of complex at base of tube

19
Q

What is a weakness of precipitation assays?

A

false-negatives

20
Q

What is an immunofluorescence assay (IFA)?

A

detect presence of antibody/antigen in serum samples

21
Q

What does the immunofluorescence assay involve?

A

binding of a specific antibody to an antigen in the sample

22
Q

What modification is done to the antibody(ies) used in the IFA assays?

A

chemically tagged with fluorescent dye = allows pathogen to be detected by fluorescent microscope

23
Q

What are the 2 types of IFA techniques?

A

direct and indirect

24
Q

What is the Direct IFA Antibody assay?

A

uses 1 antibody | detects antigens in fecal, blood, tissue samples (not serum)

25
Q

What is Indirect IFA Antibody assay?

A

uses 2 antibodies, label with fluorochrome | detects antigens in serum samples

26
Q

Define molecular techniques?

A

study of molecular basis of physiological processes that occur in microbes - such as translation and transcription

27
Q

Why are molecular techniques useful?

A

can detect slow-growing/unculturable pathogens | identify particular bacterial species present in sample via nucleic acid sequence

28
Q

What are molecular techniques commonly used as by medical professionals?

A

identify disease prognosis = allows doctors to decide on potential therapies based on specific characteristics og the pathogen

29
Q

What are the 3 molecular techniques discussed?

A

PCR | RT-PCR | electrophoretic mobility shift assay

30
Q

What is PCR?

A

polymerase chain reaction | identifies particular pathogen in a sample by a targeted nucleic acid sequence

31
Q

What is the concept behind PCR?

A

use of one DNA template strand (or many from a sample) and use of polymerase’s high efficiency to make many copies of the targeted nucleic acid

32
Q

How can you identify a particular pathogen using the PCR technique?

A

must use primer specific to a particular bacterial species

33
Q

What is PCR commonly used for?

A

identify unknown bacterial isolates and uncultivable pathogens

34
Q

What makes PCR highly usable?

A

cheap, fast, specific, uses very small sample

35
Q

What is RT-PCR?

A

reverse transcriptase-PCR | determine which genes are expressed in a particular pathogen

36
Q

What are the main steps of RT-PCR?

A

reverse transcriptase amplifies mRNA &raquo_space;> makes cDNA (complimentary DNA) &raquo_space;> PCR amplifies cDNA

37
Q

What is RT-PCR useful for?

A

determine various characteristics of a particular pathogen (ie: toxins)

38
Q

What is a weakness of RT-PCR?

A

unstable nature of RNA | RNase can degrade product

39
Q

What are the results of most immunoassays expressed as?

A

positive = presence of Ab/antigen | negative = absence of Ab/antigen

Medical Microbiology (20 decks)

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