Profiling and micro biome study Flashcards

1
Q

reasons to study the microbiome

A

eradicate harmful components - h.pylori, ulcers
treat diseases more naturally
prevent disease formation - lack of natural protection and dysbiosis

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2
Q

diseases now considered affected by the micro biome

A

psoriasis, BV, Chrons, obesity and acne

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3
Q

16S RNA sequencing

A

all bacteria have this as a part of their 30s subunit of prokaryotic ribosome

  • highly conserved and variable regions
  • identify and distinguish between taxonomic groups
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4
Q

organisms which are very closely related

A

same (hyper)variable region - newly synthesised

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5
Q

METAGENOME SEQUENCING

A
  • whole genome sequencing
  • functions and pathways
  • essential for defining the microbiome
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6
Q

what does a profiling technique need to be

A
  • able to deal with huge amounts of data generated
  • culture independent sequence analysis - small amount able to be cultured
  • reproducible and representative
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7
Q

how many genomes currently sequenced from gut microbiota

A

178 - 2phyla

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8
Q

Enterobactericae

A

gram -ve
e.coli - opportunistic
salmonella - obligate

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9
Q

WGS

A

genome characteristics, outbreak analysis, antibiotic susceptibility testing and virulence investigations

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10
Q

WGS was used to investigate

A

transmission in hospitals of vancomycin resistant E.faecilbacterium

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11
Q

WGS can provide

A

a map of the virulome

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12
Q

MALDI-TOF MS

A

matrix assisted laser desorption/ionisation tim of flight mass spectrometry

  • gold standard for microbial identification
  • fast cheap and reliable
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13
Q

MALDI TOF MS can identify

A

bacteria and fungi cultivated on agar plates or in liquid medias

  • antibiotic/myotic resistance testing
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14
Q

what does MALDI TOF rely on

A

RIA and ELISA

radio immune assay and enzyme linked immunosorbent assay

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15
Q

MALDI TOF overview

A
  • antibody + mass spec]
  • can see small proteins and peptide in forces - calibration curve tabled
  • stable isotopes
    toxin quantification is easy
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16
Q

IMS

A

imaging mass spectrometry - being developed - will be very detailed

17
Q

image mass spectrometry will be able to

A

analyse proteins, peptide, modifications, drugs and metabolites without labelling them