Objective: This study was focused on assessment and performance of conventional and nanotech assisted methods for detection of Mycobacterium tuberculosis. Tuberculosis (TB) still leftovers one of the top ten causes of death worldwide, thus this study has undertaken the use of MNP for early detection of TB.Methods: A cross-sectional studies were conducted on clinical and radiological suspected TB patients in the department of microbiology at D. Y. Patil hospital Kolhapur. All samples received in the month of August 2016 to January 2017. Total One hundred twenty-sputum samples were processed for diagnosis of TB by ZN stain, culture on L. J. Medium and real-timePCR tests.Results: In our present study, 60.83% (73) patient male and 39.17 % (47) were female patient, showing a borderline male prevalence found in our study. Eighteen percent patients were found to be in the age group (21-30 y) are mostly affected for pulmonary tuberculosis. A significant difference was seen in the percentage of differentDNA extraction methods, the fig. being conventional chloroform-phenol 66.66%, the commercial kit 80%, magnetic bead 86.66% and MNPs method 99.66% found to be significant (P<0.0001**).Conclusion: In the present study the MNP-DNA extraction techniques with NALC followed by IS6110 target amplification were found superior for diagnosis of TB. The MNP assisted extraction method showed better results in terms of quantification and sensitivity of TB PCR diagnosis, evolving nanotech assisted innovative method.

Objective: Tuberculosis is one of the leading causes of death worldwide. It is present in all the countries and among all age groups and also seen in both the genders. In investigating the early stage, identification, and rapid detection of tuberculosis, the PCR method (polymerase chain reaction) is one of the fast, safest, and reproducible new approaches. It covers all advantages such as the use of closed systems, reduced risk of carryover contamination, improved sensitivity and reproducibility, reduced turnaround time wide dynamic range of target detection, and feasibility for quantitation making it easy and reliable for the early detection of Mycobacterium tuberculosis. Methods: All samples used for the study are confirmed by conventional microscopic observation using acid-fast staining using ZN STAIN. The pulmonary sputum samples are obtained from clinical and radiological evidence of tuberculosis and these samples are selected for DNA extraction. Discussion: As per the statistical analysis using SPSS 22 version, it is found that the TB positivity rate (is 29.75%). Out of 119 positive patients (Male: 60.5% and Female: 39.5%). The average and standard deviation for CT values are 23.6 and 2.9, respectively. The 95% confidence interval of CT values for specimens is (22.7, 24.5). Average and standard deviation for CT values are equal in male patients and female patients. Age group-wise average and standard deviation for CT values are nearly equal. Results: The percentage of 3+ AFB positive grades having CT values between 20 and 24 is more than all other combinations of AFB positive grades and CT values. The average and standard deviation for CT values are 23.6 and 2.9, respectively. The 95% confidence interval of CT values for specimens is (22.7, 24.5). The average and standard deviation for CT values are equal in male patients and female patients. Conclusion: A significant improvement in specificity with high accuracy was achieved by a real-time PCR assay. Real-time PCR tests prove both a high degree of sensitivity in the upper respiratory sputum samples and for the early detection of the TB infection. The MTB real-time PCR test suitably identified the majority of the AFB positive and bacterial culture confirms TB.

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