Identification of Mycobacterial MPT-64 and ESAT-6 Proteins in Urogenital Tuberculosis Patients by Real-Time Immuno-PCR

Abstract

Aim: Diagnosis of urogenital tuberculosis (UGTB) is difficult and there is an immediate need to develop a reliable diagnostic test. Methods: A real-time immuno-PCR (RT-I-PCR) was developed to identify a cocktail of MPT-64 + ESAT-6 in both male/female UGTB patients comprising five confirmed cases, 40 clinically suspected cases and 37 non-TB controls, from whom mid-stream urine specimens were collected, while endometrial biopsies of female patients were obtained on day 1 of their menstrual cycle. Results obtained by RT-I-PCR were compared with I-PCR/ELISA and GeneXpert. Results: A wide range (500 fg/ml–10 ng/ml) of MPT-64 + ESAT-6 was detected in UGTB specimens by RT-I-PCR, although ELISA showed a narrow range (2.5–11 ng/ml). Sensitivities of 80% and 82.2% were obtained by RT-I-PCR in clinically suspected and total UGTB cases, respectively, whereas 94.6% specificity was obtained. Concurrently, RT-I-PCR revealed significantly higher (p < 0.05–0.001) sensitivity than I-PCR/ELISA and GeneXpert. Conclusion: After improving the specificity, the authors may develop RT-I-PCR into a diagnostic kit.

Genetic diversities of Mycobacterium tuberculosis complex species in Western Kenya

“…Prior to the onset of the coronavirus disease 2019 (COVID-19) pandemic, TB stood as the primary cause of mortality attributed to a single infectious agent. However, in the current context TB has become the second leading infectious cause of death, following closely behind COVID-19 [ 7 ].…”

“…It plays a pivotal role in the virulence of these mycobacteria by contributing to the evasion of host immune responses and facilitating cell invasion, as highlighted by Kamra et al . [ 7 ]. Another noteworthy protein is the antigen 85 complex (Ag85), which is conserved across various mycobacterial species, including the MTBC and some non-tuberculous mycobacteria (NTM).…”

“…This protein is actively involved in the biosynthesis of mycolic acids, crucial components of the mycobacterial cell wall, as elucidated by Khan et al . [ 7 ].…”

Diagnosis of genitourinary tuberculosis: detection of mycobacterial lipoarabinomannan and MPT-64 biomarkers within urine extracellular vesicles by nano-based immuno-PCR assay

“…In developing countries, smear microscopy is widely used but it lacks sensitivity as well as reproducibility 3 . Though culture identification of Mycobacterium tuberculosis ( Mtb ) from endometrial biopsies (EBs)/urine specimens of GUTB cases is considered to be the gold/reference standard, it generally exhibits low sensitivity on Lowenstein-Jensen (LJ) medium due to meager bacterial load within them 4 , 5 . Meanwhile, it has a long turnaround time of 4–6 weeks and needs expertise of skillful technicians 5 .…”

“…Though culture identification of Mycobacterium tuberculosis ( Mtb ) from endometrial biopsies (EBs)/urine specimens of GUTB cases is considered to be the gold/reference standard, it generally exhibits low sensitivity on Lowenstein-Jensen (LJ) medium due to meager bacterial load within them 4 , 5 . Meanwhile, it has a long turnaround time of 4–6 weeks and needs expertise of skillful technicians 5 . Histopathological examination (HPE) is an important technique to diagnose GUTB 5 , albeit HPE is not pathognomonic except for the presence of acid-fast bacteria (AFB), since it cannot differentiate between GUTB and other granulomatous diseases 6 , 7 .…”