In caseous necrotic areas of TB lesions, occasional but strong signals were detected.
The staining pattern was predominantly granular with anti-MPT64 , whereas it was predominantly diffuse with anti-BCG antibodies with a similar localization. Results of various tests were compared between the TB and the non-TB specimens. The strength of immunostaining using anti-MPT64 is that it is simple, rapid, robust, sensitive, provides clear and intense signals, makes it possible to differentiate M.
It can easily be availed in any clinical pathology laboratory. One of the most important advantages is that even very tiny specimens are suitable for processing, which is extremely important as it is very difficult to get sufficient specimens from inaccessible EPTB lesions. Thus, immunostaining with anti-MPT64 should be a diagnostic procedure in the routine evaluation protocol of EPTB cases in the surgical pathology laboratory.
Our results show that immunostaining with anti-MPT64 is much more sensitive than ZN microscopy and culture. Also, immunostaining is significantly faster compared with culture as the results are available within 1 working day. Also, this assay has a sensitivity and specificity comparable to that of nested-PCR, with the advantage of being simple, robust, and not sensitive to contamination and does not require sophisticated equipments, which make it suitable for implementation in low-income and middle-income countries with relatively modest laboratory facilities.
The most important advantage of the assay is that we could confirm granulomatous and necrotic lesions confidently not only on biopsy, but also with cytology material, which is very important in EPTB cases Table 2. We overdiagnose 1 control specimen as tuberculous lesion with anti-MPT64 immunostaining. However, misdiagnosis and overdiagnosis were more often observed with anti-BCG immunostaining. Probably, the lower specificity with anti-BCG, the only commercially available antibody for TB, due to cross-reactivity with other infectious organisms 21—23 could be the reason for the restraint of immunostaining as the routine assay for the evaluation of granulomatous lesions in the surgical pathology laboratory.
However, immunostaining with anti-MPT64 antibodies provides sharp and strong signals with a clear background compared with anti-BCG antibodies, making interpretation easier and permitting a more confident diagnosis of M. Because of the low sensitivity of the culture in pauci-bacillary EPTB, we used positive nested-PCR to define TB cases and for the validation of the immunostaining assay. All culture-positive cases were positive with nested-PCR and immunostaining. As the sensitivity and the specificity of the anti-MPT64 immunostaining assay was very similar to that of nested-PCR, we once again prove its reliability for the confirmation of TB in resource-constrained settings.
A further multicenter study with a larger number of specimens may help decide its use in clinical practice as a standalone and replacement of the ZN stain test for EPTB cases. In conclusion, our study provides the insight for the feasibility of using the technique as a standalone diagnostic technique for better management of EPTB patients in settings where culture or molecular facilities are not available.
We believe that immunostaining using anti-MPT64 for the identification of M. The authors are grateful to Dr V. Mahadik, Director of R. Gardi Medical College, for his encouragement and support during this work. You may be trying to access this site from a secured browser on the server. Please enable scripts and reload this page.
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Since October , Dr. He completed an internal medicine residency and a chief medical residency at the University of Connecticut Health Center, Farmington, and an infectious disease fellowship at Yale University School of Medicine. Address correspondence to Holenarasipur R. Vikram, M. Shea Blvd. Reprints are not available from the authors. Centers for Disease Control and Prevention. Consensus statement. Global burden of tuberculosis: estimated incidence, prevalence, and mortality by country. Extrapulmonary tuberculosis in the United States. Am Rev Respir Dis. Tuberculosis in patients with the acquired immunodeficiency syndrome.
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