NEW METHODS FOR DIAGNOSIS OF TUBECRCULOSIS

Abstract

This work was done on 40 patients diagnosed clinically to have tuberculosis [29 males and I I females] whose age ranged froin .I 2 to 65 years. Eleven patients received the nine months course of treatment and still positive when ex ami ned by acid -fast microscopy and the 29 were defaulter patients.

A.ll patients were subjected to:

Detailed history, full clinical examination and chest X-ray. Sputuin samples were subjected to decontamination, liquefaction and concentration procedures using N-acetyl-L-cystine NaOH 4% method.

Decontaminated, concentrated samples were subjected to the following:

-Routine Microbiology tests:

A-Direct staining using: Zeihl Neelsen stain & Auramine Rhodamine fluorescent stain.

B- Culture of specimen on: Conventional Lowenstain 1enn media &

Mycobacterial Growth Indicator Tube (MG.IT) media

C-Drug susceptibility test by: Proportion agar dilution test.

-Molecular biology techniqtles:

A- Extraction of DNA using Mini-bead agitation inethod,

B- Polymerase Chain Reaction amplifying a DNA target segment: including: Multiplex PCR for IS6110 & JS/245 fot detection of M tuberculosis and M aviwi1, 65 kDa HSP and 16S rRNA for detection of Mycobacterium and oxyR gene for differentiation between

Mtuberculosis and M.bovis.