Rapid Microbial Methods - Applied Microbiology - Lecture Slides, Slides of Microbiology

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Recent

Advances

in

Rapid

Microbial

Identification

and

Characterization

Techniques

Traditional

Microbial

Methods

Robert

Koch

and

Pure

Cultures

His study of infectious diseases lead to the rise of pure cultures Started using potato slices Gelatin: solution for potato slices

Traditional

Microbial

Methods

Richard

Petri

Double sided clear petri plates Easy to stack, sterilize, view colonies

Traditional

Microbial

Methods

Martinus

Beijerinck

and

Enrichment

Culture

Manipulated incubation and nutrient conditions of natural samples Was able to isolate and identify the first pure cultures of soil and aquatic microorganisms

Lactobacillus species - green algae Was the first to describe a virus - Tobacco mosaic virus

Microscopy

and

Staining

Techniques

Viability staining Fluorescent staining using

DAPI

diamido

phenylindole)

What

Are

Rapid

Microbial

Methods

(RMM)?

Technologies

that

allow

users

to

get

microbial

test

results

faster

than

traditional

methods

Can

be

used

for

qualitative,

quantitative

or

identification Common

Technologies

Used

**Just list the ones we are going to talk about

i will** mention more but say “our focus is...”

Immunoassays

Advantages

Easy to use

Cost effective - Fast detection

hour to

minutes)

Disadvantages

Low sensitivity

Better to use heavily contaminated waters and larger size pathogens - Not successful in detecting enteroviruses

ELISA:

Enzyme

Linked

ImmunoSorbant

Assay

Purpose:

to

determine

if

a

particular

protein

or

target Microtiter

Plates

Used

Forms

Direct - Indirect

Indirect

Elisa

Used

to

detect

presence

of

specific

antibody

in

specimen

 Initial step where Antigen is bound to surface of plate  Sample added  Final step where substrate for enzyme is added  Colour change indicates sample contains antibody that reacts against antigen (Left well)

Immunoliposome

Technology

Involves

things

Liposomes with encapsulated dyes

Antibodies to antigen of microbe tagged outside

Immunostrip

Kit

by

Agida

Inc.

Respiring

Microbe

Method:

ATP

Production

Excellent

way

of

measuring

presence

of

microbes

in

recreational

contaminated

waters

Advantage

fast, simple

Disadvantage

Careful calibration for sample needed

LuminUltra

Technology

Fast - Portable - Complete

Benefits

and

Drawbacks

of

RMM

Benefits

Faster time - Allows companies to release finished products to the market in less time - Ability to identify, contain and recover from contaminant events - Reduce worker subjectivity - Take guesswork out of plating and waiting - Less training - Ease of use - Higher specificity/sensitivity Drawbacks - Can’t replace traditional methods - Cost per test high - Complex technology

Saponification

of

cellular

lipids

and

Fatty

acid

methylation

Triglyceride Methanol Glycerol FAMEs

Saponification

of

cellular

lipids

and

Fatty

acid

methylation

Triglyceride Methanol Glycerol FAMEs