Serology, Molecular, and Proteomic Methods
>
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Diagnostic tests
• specific - identify target (minimize false-positives) - > Polymerase chain reaction (PCR)
• sensitive - detect in low abundance (minimize false- • sensitive - can detect single pathogen in
negatives) sample
• accurate - both • facilitates gene sequencing for genetic
> antibody functions disorders, diagnosing infections
> MALDI-TOF MS
• activate complement cascade, neutralization,
opsonization, agglutination, antibody dependent cell • matrix assisted labor desorption/ionization
mediated cytotoxicity time of flight mass spectrometry
• antigen-binding site of antibodies • fast (2 days)
- highly specific, strong noncovalent interactions • can't distinguish between some pathogens
>
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immunological diagnostic tests
• biochemical tests - identify bacteria responsible for
infection
- can take more than 24 hrs, pathogen must be
culturable, can't identify noncellular
• typically involve serology
• determine if patient has certain antigens/antibodies in
blood
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agglutination tests
• antibodies have 2 antigen binding sites - can bind
antigens into clump
• seen in blood agglutination tests
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plaque reduction neutralization test (PRNT)
• patients serum extracted and diluted, petri plates,
incubated
• positive: neutralizing antibodies present in sample,
antibodies bind to virus and neutralize them
> enzyme linked immunosorbent assays (ELISAs)
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• versatile and rapid
• reporter enzyme attached to monoclonal detecting
antibody
>
- lateral flow (Covid and others)
• capillary flow down test into test lines
= immunofluorescence microscopy
• fluorescent tagged antibodies to recognize specific
antigen in sample
=
immunofluorescence assays (IFAs)
• detects antigens or antibodies in patient sample
• detection antibody linked to fluorescent tag
• eliminates need to add substrate for detection step
(ELISA)
>
- interferon gamma release assays (IGRAs)
• fast and reliable, detect TB
• measure patient T cell response to mycobacterium
tuberculosis antigens
- with TB, T cells release more interferon gamma
>
-
Diagnostic tests
• specific - identify target (minimize false-positives) - > Polymerase chain reaction (PCR)
• sensitive - detect in low abundance (minimize false- • sensitive - can detect single pathogen in
negatives) sample
• accurate - both • facilitates gene sequencing for genetic
> antibody functions disorders, diagnosing infections
> MALDI-TOF MS
• activate complement cascade, neutralization,
opsonization, agglutination, antibody dependent cell • matrix assisted labor desorption/ionization
mediated cytotoxicity time of flight mass spectrometry
• antigen-binding site of antibodies • fast (2 days)
- highly specific, strong noncovalent interactions • can't distinguish between some pathogens
>
-
immunological diagnostic tests
• biochemical tests - identify bacteria responsible for
infection
- can take more than 24 hrs, pathogen must be
culturable, can't identify noncellular
• typically involve serology
• determine if patient has certain antigens/antibodies in
blood
-
agglutination tests
• antibodies have 2 antigen binding sites - can bind
antigens into clump
• seen in blood agglutination tests
-
plaque reduction neutralization test (PRNT)
• patients serum extracted and diluted, petri plates,
incubated
• positive: neutralizing antibodies present in sample,
antibodies bind to virus and neutralize them
> enzyme linked immunosorbent assays (ELISAs)
-
• versatile and rapid
• reporter enzyme attached to monoclonal detecting
antibody
>
- lateral flow (Covid and others)
• capillary flow down test into test lines
= immunofluorescence microscopy
• fluorescent tagged antibodies to recognize specific
antigen in sample
=
immunofluorescence assays (IFAs)
• detects antigens or antibodies in patient sample
• detection antibody linked to fluorescent tag
• eliminates need to add substrate for detection step
(ELISA)
>
- interferon gamma release assays (IGRAs)
• fast and reliable, detect TB
• measure patient T cell response to mycobacterium
tuberculosis antigens
- with TB, T cells release more interferon gamma
