Volume 14 Supplement 3
Use of multi-primer / multi-gene PCR method for the detection of Mycobacterium tuberculosis among female genital tuberculosis patients in India
© Bhanothu et al; licensee BioMed Central Ltd. 2014
Published: 27 May 2014
Female genital tuberculosis (FGTB) is a silent disease that evidences itself only when it is investigated. Demonstration of the etiologic agent by H & E staining or Z-N staining for acid fast bacilli, smear microscopy, and culture of sputum and other body fluid specimens were often less specific.
A prospective case-control study was undertaken. A total of 302 endo-ovarian tissue biopsies collected from 202 infertile women highly suspected of having genital tuberculosis on laparoscopic examination and from 100 control women of reproductive age. All specimens tested by conventional/phenotypic methods were later compared with multi-gene/ multi-primer PCR method using four sets of primers for detection of Mycobacterium tuberculosis (MTB) in a single tube-single step reaction.
The conventional methods showed 99% to 100% specificity with a low sensitivity, ranging from 21.78% to 42.08% while H & E staining showed a sensitivity of 51.48%. Multi-gene PCR method was found to have a much higher sensitivity of 70.29% with MTB64 gene, 86.63% with 19kDa antigen gene at species and TRC4 element at regional MTB complex level and 88.12% with MPT59 α-antigen/32kDa protein gene at genus level. The specificity of multi gene/multi primer PCR was 100%.
Multi-gene PCR was found to be a powerful technique for diagnosis and differentiation of mycobacterial infection among endo-ovarian tissue biopsies taken from infertile patients with FGTB. We suggest site specific sampling and amplification of the 19kDa antigen gene in combination with TRC4 element as a successful multi-gene/ multi-primer PCR method for the diagnosis of FGTB.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.