PURPOSE: Diagnosis of extrapulmonary tuberculosis (EPTB) poses serious challenges. A careful selection of appropriate gene targets is essential for designing a multiplex-polymerase chain reaction (M-PCR) assay. MATERIALS AND METHODS: We compared several gene targets of Mycobacterium tuberculosis, including IS6110, devR, and genes encoding MPB-64 (mpb64), 38kDa (pstS1), 65kDa (hsp65), 30kDa (fbpB), ESAT-6 (esat6), and CFP-10 (cfp10) proteins, using PCR assays on 105 EPTB specimens. From these data, we chose the two best gene targets to design an M-PCR. RESULTS: Among all gene targets tested, mpb64 showed the highest sensitivity (84% in confirmed cases and 77.5% in clinically suspected cases), followed by IS6110, hsp65, 38kDa, 30kDa, esat6, cfp10, and devR. We used mpb64+IS6110 for designing an M-PCR assay. Our M-PCR assay demonstrated a high sensitivity of 96% in confirmed EPTB cases and 88.75% in clinically suspected EPTB cases with a high specificity of 100%, taking clinical diagnosis as the gold standard. CONCLUSION: These M-PCR results along with the clinical findings may facilitate an early diagnosis of EPTB patients and clinical management of disease.
Bacteriological Techniques/methods ; DNA Transposable Elements/genetics ; DNA, Bacterial/analysis/genetics ; Early Diagnosis ; Female ; Gene Amplification ; Humans ; Male ; Multiplex Polymerase Chain Reaction/*methods ; Mycobacterium tuberculosis/genetics/*isolation & purification ; Polymerase Chain Reaction/*methods/standards ; Sensitivity and Specificity ; Tuberculosis/*diagnosis

Objective: With advancements in cardiac surgical interventions during infancy and childhood, the incidence of maternal congenital heart disease (CHD) is increasing. This retrospective study compared fetal and cardiac outcomes in women with and without CHD, along with a sub-analysis between cyanotic versus non-cyanotic defects and operated versus non-operated cases. Methods: A 10-year data were retrospectively collected from pregnant women with CHD and a 1:1 ratio of pregnant women without any heart disease. Adverse fetal and cardiac outcomes were noted in both groups. Statistical significance was set at P<0.05. Results: A total of 86 pregnant women with CHD were studied, with atrial septal defects (29.06%) being the most common. Out of 86 participants, 27 (31.39%) had cyanotic CHD. Around 55% of cases were already operated on for their cardiac defects. Among cardiovascular complications, 5.8% suffered from heart failure, 7.0% had pulmonary arterial hypertension, 8.1% presented in New York Heart Association functional class IV, 9.3% had a need for intensive care unit admission, and one experienced maternal mortality. Adverse fetal outcomes, including operative vaginal delivery, mean duration of hospital stay, fetal growth restriction, preterm birth (<37 weeks), low birth weight (<2,500 g), 5-minute APGAR score <7, and neonatal intensive care unit admissions, were significantly higher in women with CHD than in women without heart disease. Conclusion Women with CHD have a higher risk of adverse fetal and cardiac outcomes. The outcome can be improved with proper pre-conceptional optimization of the cardiac condition, good antenatal care, and multidisciplinary team management.