Introduction: Non-tuberculous Mycobacteria (NTM) pulmonary disease is often unrecognised or misdiagnosed as Mycobacterium tuberculosis (MTB), Multi-Drug Resistant Tuberculosis (MDRTB), because of similar clinical presentation in counties with high burden of Tuberculosis (TB) including India. In India due to lack of awareness among clinicians and lack of laboratory facilities to diagnose these infections, its prevalence is largely unknown. Aim: To evaluate the efficacy of identification of NTM species by Matrix Assisted Laser Desorption Ionisation Time of Flight (MALDI-TOF) and Heat Shock Protein (hSP65) gene sequencing and to determine their Antimicrobial Susceptibility Testing (AST). Materials and Methods: All the clinical specimens from pulmonary and extra-pulmonary TB suspects at Nizam’s Institute of Medical Sciences, Hyderabad, Telangana, India, over a period of one year i.e., from June 2017 to May 2018 were included in the study. The specimens were subjected to microscopy, culture and GeneXpert. The NTMs isolated in the culture were further characterised genotypically by MALDI-TOF and hsp65 gene sequencing. The identified NTM isolates were subjected to AST. All the methods were followed as per the standard protocols. Data was analysed using SPSS 25. Results: A total of 1085 samples were processed out of which Mycobacteria was detected in 201 cases (18.5%). Among the culture positives, MTB complex was detected in 146 cases (13.5%) and NTM in 55 (5.06%). Mycobacterium abscessus was the predominant isolate. The most common co-morbidities were bronchiectasis and Chronic Obstructive Pulmonary Disease (COPD). Linezolid, clarithromycin, moxifloxacin and amikacin showed high sensitivity. Conclusion: Molecular assay helps in rapid identification which can lead to targeted therapy and can thus combat antimicrobial resistance. The MALDI-TOF and hsp65 gene sequencing also offers quick results at a low cost and is easy to perform hence it can be considered as an alternate diagnostic tool for identification.

Article Subject: Medical Microbiology 10.30699/ijmm.13.3.172 Background and Aims: Tuberculosis kills more than 1 million people every year, most of them in low-income and middle-income countries. An understanding of the trends in tuberculosis incidence, prevalence, and mortality s crucial to track the success of tuberculosis control programs. Microbiological diagnosis of diseases caused by Mycobacteria should be fast and effective to prevent contagions and optimize the management of infections. Materials and Methods: A total of 1412 clinical pulmonary and extra pulmonary specimens were studied from January 2017 to December 2017 at Nizam's Institute of Medical Sciences, Hyderabad. All specimens were processed according to standard operating procedures. All the specimens were subjected to microscopy, culture, GeneXpert. Results: Among 1412 samples received 813 were males (57.6%) and 599 females (42.4%). Among these 818 (57.9%) were pulmonary samples and 594 extra pulmonary samples. Mycobacterium prevalence was (21.6%) out of which Mycobacterium tuberculosis was found in 18.3% and Non tuberculous Mycobacteria (3.25%). The contamination rate was 2.6% (37 out of 1412). Among the positives, the most common affected age group was 21-30 yrs (22.2%). About 64 (4.53%) were smear positive. A total of 200 isolates (14.16%) were recovered by at least one culture LJ medium or BacT Alert 3D system. MTB was recovered in 216 (15.29%) by GeneXpert. MDRTB was detected in 8 (3.7%) by GeneXpert. Conclusion: Mycobacterium tuberculosis complex is responsible for immense worldwide morbidity and mortality. Delays in diagnosis may postpone administration of appropriate treatment and be detrimental to patient outcomes. Since traditional culture methods are slow, newer molecular techniques allow more rapid and sensitive laboratory diagnosis of tuberculosis.

Introduction: Intestinal coccidian parasites known to cause opportunistic infections have increased since the past years and are often indistinguishable from other forms of community- acquired diarrhoea, which suggests a need for proper and rapid diagnostic techniques to recover and identify the organism. Aim: To investigate if Auramine Rhodamine (AR) staining is a sensitive and time conserving staining technique in contrast to modified acid fast stain. Materials and Methods: A descriptive observational study was conducted at Nizam’s Institute of Medical Sciences, Hyderabad, India, over a period of three months (i.e., from July to September 2019). A total of 100 stool samples received by the Department of Microbiology were subjected to macroscopic and microscopic examination by saline mount, iodine mount, kinyoun acid fast stain, modified AR staining. Results: In the present study the prevalence of coccidian infections were observed to be 6%. Coccidian parasites reported were Cryptosporidium species and Cystoisospora species immunocompromised patients were found to be more prone to infections with coccidian parasites. The AR stain showed 100% agreement with modified acid fast stain. Conclusion: This study concluded that the AR stain is a rapid and better stain than kinyoun acid-fast stain for the detection of coccidian parasites.