Research Article Open Access Faridi et al., J Med Microb Diagn 2018, 7:4 DOI: 10.4172/2161-0703.1000285 Research Article Open Access Journal of Medical Microbiology & Diagnosis ISSN: 2161-0703 J o u r n a l o f M e d i c a l M i c r o b i o l o g y & D i a g n o s i s Volume 7 • Issue 4 • 1000285 J Med Microb Diagn, an open access journal ISSN: 2161-0703 *Corresponding author: Dr. Maryam Faridi, Department of Microbiology, J.N. Medical College, AMU, Aligarh, Uttar Pradesh, India, Tel: 0571 522 8770; E-mail: maryamfaridi0@gmail.com Received September 18, 2018; Accepted October 22, 2018; Published November 05, 2018 Citation: Faridi M, Shukla I, Fatima N, Varshney S, Shameem M (2018) Prevalence of Primary Pulmonary Multi-Drug Resistant Tuberculosis in and around Aligarh Region. J Med Microb Diagn 7: 285. doi:10.4172/2161-0703.1000285 Copyright: © 2018 Faridi M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Abstract Tuberculosis (TB) is one of the most ancient diseases of mankind and has co-evolved with humans for many thousands of years or perhaps for several million years. M. tuberculosis strains that are resistant to the two most potent anti-TB drugs Isoniazid and Rifampicin, are termed as multidrug-resistant TB (MDR-TB) strains. Drug resistance is broadly classifed as primary and acquired. Drug resistance in a patient who has never received anti-TB treatment previously or has taken treatment for less than a month is termed as primary resistance. Acquired resistance is the resistance which arises as a result of specifc previous treatment. This study was aimed to determine the prevalence of primary MDR-TB in and around Aligarh region by molecular diagnostic method of Line probe assay (LPA). This two year study was carried out in culture and DST Laboratory (RNTCP certifed), Department of Microbiology, J.N. Medical College AMU, Aligarh on the sputum samples received of the primary pulmonary tuberculosis suspected patients (according to PMDT guidelines) from the outpatient and inpatient departments of the hospital and from various tuberculosis units in and around Aligarh region from October 2015 to October 2017. Sputum samples were collected from suspected cases of primary pulmonary TB. These samples were subjected to routine microscopy and culture on LJ medium to detect Mycobacterium tuberculosis. Positive cases were subjected to drug sensitivity test by GenoType MTBDRplus Assay. Out of the total 514 samples collected, 265 (51.56%) samples and 326 (63.43%) samples were positive by ZN microscopy and fuorescent microscopy respectively. 312 (60.70%) samples were positive on culture on LJ medium. Total 326 samples which were positive on fuorescent microscopy were subjected to LPA and 47 (9.14%) samples were resistant to both rifampicin and isoniazid, 21 (4.08%) samples were rifampicin mono-resistant and 31 (6.03%) samples were isoniazid mono-resistant. Prevalence of Primary Pulmonary Multi-Drug Resistant Tuberculosis in and around Aligarh Region Maryam Faridi 1* , Indu Shukla 1 , Nazish Fatima 1 , Sumit Varshney 1 and Mohammad Shameem 2 1 Department of Microbiology, J.N. Medical College, AMU, Aligarh, Uttar Pradesh, India 2 Department of TB and Respiratory Diseases, J.N. Medical College, AMU, Aligarh, Uttar Pradesh, India Keywords: Tuberculosis; Drug resistance; Microscopy Introduction Tuberculosis is an infectious bacterial disease caused by Mycobacterium tuberculosis which most commonly afects the lungs. It is transmitted from person to person via droplets from the throat and lungs of people with active pulmonary disease [1]. TB is a serious global public health threat. TB is the ninth leading cause of death worldwide and the leading cause from a single infectious agent, ranking above HIV/AIDS [2]. In 2016, there were an estimated 1.3 million TB deaths among HIV negative and an additional 374 000 deaths among HIV- positive people [2]. Globally in 2016, an estimated 4.1% of new cases and 19% of previously treated cases had Multidrug resistant TB [2]. Rapid identifcation is important for efective treatment and control of MDR-TB. Conventional methods of drug susceptibility testing (DST) include solid media-based methods such as the proportion, absolute concentration, and resistance ratio methods. Tese can take up to 12 weeks to produce defnitive results, leading to prolonged infectiousness [3]. Liquid media-based tests are more rapid, but also costlier and require sophisticated laboratories and trained personnel [3]. Molecular LPA permit rapid diagnosis of TB, isoniazid and rifampin resistance, and clinically relevant non-M. tuberculosis mycobacteria. In LPA assays, DNA or RNA is isolated from culture or direct (i.e., sputum) respiratory samples and then amplifed and reverse hybridized onto a nitrocellulose strip with immobilized probes for diferent mycobacteria or for mutations that confer resistance. Tese strips can be quickly interpreted using a template, with the entire testing process taking a day or even less. Te GenoType MTBDRplus (Hain Lifesciences, Nehren, Germany) identifes rifampin and isoniazid resistance by detecting the most common mutations of the rpoB gene and the katG and inhA genes, respectively. Materials and Methods Sputum samples from 514 (n=514) suspected new pulmonary TB patients were collected and subjected to ZN microscopy and fuorescent microscopy and cultured on LJ media. Sputum positive samples were tested by LPA for the presence of M. tuberculosis complex and resistance to isoniazid and rifampicin. LPA Te GenoType MTBDRplus LPA was performed according to the manufacturer’s (Hain Life-science, Nehren, Germany) instructions. Tree steps for LPA test include DNA extraction, multiplex polymerase chain reaction (PCR) amplifcation and reverse hybridization. Tese steps were carried out in three separate rooms with restricted access and unidirectional workfow. LPA strips were observed and read for the presence of TUB band, amplifcation control band and conjugation control band and absence of any wild type (WT) band or presence of any mutation (MUT) band. Te results were then interpreted as sensitive or resistant to any particular drug.