AFB Culture

Shilpa Kathuria
060201353

Specimen collection
Specimen collection bottle.

Culture Methods

Conventional & Rapid

Conventional methods
Detects infection at a conc of 10-100 bacilli/ml Sensitivity 80-85% Specificity 98% Optimum temp: 37C High humidity pH: 6.47 Under 5% CO2 for Middlebrook media. M.Tb grow very slowly (generation time: 1415 hrs) and Visible colonies seen in 4- 6 wks.

3 types
Egg based - Lwenstein-Jensen, Petragnani, Dorset egg medium. Agar based Middlebrooks 7H10, Middlebrooks 7H11. Liquid Middlebrooks 7H9, Dubos.

Colony morphology
Solid media dry, buff coloured, raised colonies with a wrinkled surface. Liquid media colonies grow as surface pellicle due to hydrophobic nature of bacterial cell wall. virulent strains grow as serpentine cords non virulent strains have a dispersed growth.

L J medium colonies formed

Septi-chek AFB
Biphasic culture approach Combines broth & solid media into a single device. Requires 3 weeks of incubation Simultaneous detection of Mtb, other respiratory pathogens & even contaminants.

 Contains Middlebrooks 7H9 broth in CO2 enriched culture bottle & a peddle with agar media One side of peddle covered with Middlebrooks 7H11 and other side contains Middlebrooks 7H11 with NAP.

Rapid
Use of broth-based culture for isolation, determination of species and form a conclusive diagnosis in very short time period. 2 types BACTEC Radiometric method. MGIT Mycobacterial growth indicator tubes.

Bactec 460TB System

Detection in 4-8 days. Differentiation in 3-6 days. Susceptibility in 4-12 days.

BACTEC
Culture medium Middlebrooks 7H12 broth containing 14C labelled palmitic acid. This palmitic acid is metabolized by the M.Tb to release 14CO2 which can be detected by bactec 460 instrument and measured in terms of growth index.

MGIT
Consists of round bottom tubes These tubes contain middlebrooks 7H9 medium which has an oxygen sensitive flourescent sensor at the bottom. When mycobacteria grow, they deplete the dissolved oxygen in the broth & allow the indicator to fluoresce brightly in a 365nm UV light. Diagnosis made in 10 12 days. Used more frequently than bactec since it has a cheaper detection instrument.

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