IOSR Journal of Dental and Medical Sciences (IOSR-JDMS) e-ISSN: 2279-0853, p-ISSN: 2279-0861.Volume 13, Issue 11 Ver. IV (Nov. 2014), PP 50-53 www.iosrjournals.org www.iosrjournals.org 50 | Page Rapid detection of Mycobacterium tuberculosis and Rifampicin Resistance in extra pulmonary samples using Gene Xpert MTB/RIF assay Ahmad Naeem Sajed 1 , Sana khan 2 , Aneeqa Shamshad Butt 2 , Abid Srwar 3 , Dr. Raheela Akhtar 4 , Dr. Imran Ahmad 4 , Muhammad Naeem Mukhtar 1 , Dr. Sadaf Imran 4 , Dr. Hamid Akbar 4 , Shan Ali 3 , Muhammad Usman 3 1 Emergency Services Academy Rescue 1122 Lahore-Pakistan 2 Pathology Department Allama Iqbal Medical College Lahore-Pakistan 3 Institute of Molecular Biology and Biotechnology the University of Lahore-Pakistan 4 University of Veterinary and Animal Sciences Lahore-Pakistan Abstract: Tuberculosis (TB) is a major health problem in Pakistan. The World Health Organization has recently endorsed the Gene Xpert MTB/RIF assay for rapid detection of smear negative and multidrug resistance tuberculosis. A total of 100 extra pulmonary samples were processed, which included 60 pus, 19 pleural fluids, 16 ascetic fluids and 5 CSF. Out of these 37% patients were Gene Xpert MTB/RIF Assay positive, 17% were LJ culture positive and 12 % were Zn smear positive. MTB was detected in 31 out of 60 (51.7%) Pus samples, 3 out of 19 (15.8%) Pleural fluid samples, 1 out of 16 (6.3%) Ascitic fluid samples and 2 out of 5 (40.0%) CSF samples. In this study we found that Gene Xpert MTB/RIF assay is rapid method for diagnosis of extra pulmonary tuberculosis as compared to conventional methods. Because of its simplicity, rapidity and sensitivity, this seems to be a very gorgeous tool for diagnosis of extra pulmonary tuberculosis from clinical samples. Keywords: Extra pulmonary tuberculosis, Mycobacterium tuberculosis, Gene Xpert MTB/RIF assay I. Introduction Tuberculosis is the commonest infectious disease worldwide caused by Mycobacterium tuberculosis. According to WHO there were 8.6 million new TB cases in 2012 and even 1.3 million TB deaths. 1 In the under- developed countries 95% of infections occur due to pitiable diagnostic and treatment facilities. It is estimated that approximately 70 million people will die from tuberculosis within next 20 years and it is because of inadequate measures for the TB control 2 . Tuberculosis is typically divided into two types, pulmonary which is more widespread and extra pulmonary which rivet 15% cases. Extra pulmonary tuberculosis (EPTB) can involve any organ in the body. Extra pulmonary infections with members of the Mycobacterium tuberculosis complex (MTBC) have high morbidity and mortality because of lack of good diagnostic methods. Diagnosis is often difficult to establish due to low number of bacteria and collection of extra pulmonary samples is not easy. A definitive diagnosis of mycobacterium infection depends on detection of the Mycobacterium Tuberculosis in extra pulmonary samples 3 . The frequently atypical clinical presentation simulating other inflammatory and neo-plastic conditions results in a delay or deprivation of treatment which is the major challenge in the control of EPTB. Radiography provides useful information in the diagnosis of EPTB. Therefore, a high index of suspicion is necessary to make an early diagnosis and quite often, more than one procedure is necessary for the confirmation of the diagnosis. In lower-income countries, the lack of diagnostic infrastructure substantially aggravates the problem. Now attention has been devoted to latest nucleic acid amplification diagnostic systems due to their speed, sensitivity and specificity. 4 The World Health organization has recommended the use of gene Xpert for rapid detection of MTB in extra pulmonary samples. The gene Xpert MTB/RIF assay detects DNA sequences specific for Mycobacterium tuberculosis and Rifampicin resistance by polymerase chain reaction 5 . It is based on the Cepheid Gene Xpert system, a platform for rapid and simple-to-use. The Xpert MTB/RIF purifies and concentrates the Mycobacterium tuberculosis bacilli from extra pulmonary samples, isolates genomic material and amplifies the genomic DNA by PCR. Results are obtained in 90 minutes from extra pulmonary samples, with minimal biohazards. Further minimal technical training is required to operate the instrument. These features make it an important tool for extra pulmonary samples 6 .