CGMBS Ch. 11 Detection And Identification Of Micro
CGMBS ch. 11 Detection and Identification of Micro
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CGMBS ch. 11 Detection and Identification of Micro
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CGMBS Ch. 11 Detection And Identification Of Micro
Sequencing of the DNA region encoding 16S rRNA is performed to determine -
️️the evolutionary and genetic relatedness of microorganisms and has driven
changes in microorganism nomenclature
Homologous extrinsic control - ️️target-derived control with a non-target-derived
sequence insert;...
CGMBS ch. 11 Detection and Identification of Micro
CGMBS ch. 11 Detection and Identification of Micro
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CGMBS ch. 11 Detection and
Identification of Microorganisms
Sequencing of the DNA region encoding 16S rRNA is performed to determine -
✔️✔️the evolutionary and genetic relatedness of microorganisms and has driven
changes in microorganism nomenclature
Homologous extrinsic control - ✔️✔️target-derived control with a non-target-derived
sequence insert; control is added to every sample after nucleic acid extraction and
before amplification
Smallpox must be handled only in approved laboratories - ✔️✔️level 4 containment
When processing whole blood, why is it important to remove homoglobin - ✔️✔️can
inhibit DNA polymerase and prevent amplification of nucleic acid resulting in false-
negative PCR result
Inhibitors or DNA pol. in urine - ✔️✔️-nitrate
-crystals
-hemoglobin
-beta-human chorionic gonadotropin
amplification control - ✔️✔️rule out false-negatives; detects target that is always
present
Housekeeping genes in prokaryotes - ✔️✔️groEL, rpoB, recA, gyrB
Housekeeping genes in eukaryotes - ✔️✔️B-actin, glyceraldehyde-3-phosphate,
interferon-y, extrinsic homologous control, human mitochondril DNA, peptidylprolyl
isomerase A
Salmonella typhi specimen collection - ✔️✔️initially present in peripheral bloo but not in
urine or stool util at least 2 weeks after infection
Heterologous extrinsic controls - ✔️✔️non-target-derived controls that are added to
every sample before nucleic acid extraction; second set of primers must also be added
to the reaction for the conrol to be amplified
Heterologous intrinsic ontrols - ✔️✔️nontarget sequences natrually present in the
sample, such as eukaryotic genes in a test for microorganisms
, Reasons for false-negative results on a sample - ✔️✔️1. organism may be present, but
nucleic acid was degraded during collection, transport, and or extraction
2. amplification procedures may be inhibited by substances present in the specimen
Microbiological identification and epidemiology - ✔️✔️mass spectrometry of microbial
proteins
Bordetella pertussis - ✔️✔️-pathogen of upper respiratory tract that causes whooping
cough
-transmitted via direct contact with infected respiratory secretions
-detection by qPCR targeting IS481 and IS1001
Legionella pneumophila - ✔️✔️-cause of Legionnaires' disease
-infection of lower respiratory tract
-found in water
-PCR targets the macrophage infectivity potentiator(mip) gene and 16S and 5S rRNA
genes
M. tuberculosis - ✔️✔️-cause of respiratory tract infections, significant levels of
mobidity and mortality
-4.4 million bp; 4k genes
-fluorochrome stain
-10^4 organisms/mL required to see mycobacteria in a smear
-Cultures are more sensitive than smears
-PCR tests are specific to species; IS6110 and 16S rRNA for M. tuberculosis from fresh,
frozen, or fixed tissue
-qPCR with primers and probes targeting rRNA internal transcribed spacer (ITS)
elements
Mycoplasma pneumoniae - ✔️✔️-multilocus VNTR, multilocus sequence typing,
MALDI-TOF MS
Streptococcus pneumoniae - ✔️✔️-common cause of bacteremia, sepsis, otitis media,
meningitis
-positive PCR asay is questionable b/c significant portion of the population ie children is
colonized with the organism, and PCR cannot discern between colonization and
infection
Nsisseria gonorrhoeae
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