Mycobacterial load is limiting in samples of pleural tuberculosis, hence its isolation and identification is demanding. Direct examination of pleural fluids by Ziehl-Neelsen staining requires bacillary densities of 10,000/ml, whereas for isolation by culture 10-100 viable bacilli are Capsaicin needed. Detection of mycobacteria in suspected cases of pleural effusion has been shown to be augmented by the inclusion of sputum. Variable sensitivity ranging from 3.5 to 100% of isolation by culture from sputum collected from tuberculous pleural effusion patients has been reported,. However in general the isolation of Mtb from pleural fluid has been lower compared to sputum,, with the exception of the reports by Seibert et al & Epstein et al. Similarly the direct AFB smear microscopy of pleural fluid samples derived from pleural effusion TB patients was lower as compared to sputum collected from these patients,. AFB positivity of sputum smears of pleural effusion patients ranged from 1.7�C62.5%. Besides inclusion of an assortment of samples from an individual patient, attempts have been made to use rapid reliable DNA amplification techniques for efficient diagnosis of tuberculosis. The sensitivity of the different PCR assay has been reported to be ranged from 43.4�C73.8%. In the present study, we have evaluated the inclusion of sputum along with pleural fluid and the usefulness of the direct identification and detection of Mtb using an in house N-PCR assay. It can be seen that the detection of mycobacteria in pleural fluid and sputum samples varied. Ten pleural fluid samples were positive, whereas the sputum samples of these patients collected in tandem were negative. Similarly eleven sputum samples were positive whereas the pleural fluid samples of these patients were negative by the N-PCR assay. The inclusion of sputum samples of clinically diagnosed pleural TB patients increased the total number of pleural TB patients Pepstatin A detected from 30 to 41. The sensitivity of the assay increased from 51.7 to 70.6%,. Leptospirosis is a zoonotic disease caused by pathogenic bacteria of the genus Leptospira, which are transmitted directly or indirectly from animals to humans.