Molecular Characterization of katG and inhA Mutations by Genotype MTBDRplus Line Probe Assay To Guide Isoniazid and Ethionamide Use for Drug-Resistant Tuberculosis.

Introduction Drug-resistant tuberculosis (TB) continues to be a global health threat in all its forms. Significant resistance has been observed against isoniazid (INH), one of the most important therapeutic options for treating TB. Molecular testing methods such as line probe assay (LPA) provide rapid diagnosis and early management. Mutations in different genes can be detected, which indicate INH and ethionamide (ETH) drug resistance. We aimed to determine the frequency of these mutations in katG and inhA genes by LPA to guide INH and ETH use for drug-resistant TB. Materials and methods Two consecutive sputum samples were collected from each patient, followed by decontamination by N­acetyl­L­cysteine and sodium hydroxide method. LPA was performed on the decontaminated samples by GenoType MTBDRplus, and the strips were analyzed. Results Out of the 3,398 smear-positive samples tested by LPA, valid results were found in 3,085 (90.79%) samples. Of the 3,085 samples, INH resistance was seen in 295 samples (9.56%), of which mono INH resistance was in 204 samples, and 91 were multidrug resistant. katG S315T was the most common mutation responsible for high-level INH resistance. At the same time, inhA c15t was the most common mutation associated with low-level INH resistance and ETH cross-resistance. The average turnaround time for the processing and reporting of samples was five days. Conclusions The high burden of INH resistance is alarming and can be a major obstacle to TB elimination. Although molecular methods have reduced the reporting time leading to early management of the patients still, a large knowledge gap persists.

Speciation, Biofilm Formation and Antifungal Susceptibility of Candida Isolates from Clinically Diagnosed Patient of UTI in a Tertiary Care Hospital.

INTRODUCTION: The incidence of the urinary tract infections caused by Candida species, are becoming more common. Recently, an increase in the incidence of infection caused by fungi especially non albicans candida species (NAC) has been reported. Several virulence factors like biofilm formation, toxin production and presence of adhesins contribute to its pathogenesis. OBJECTIVES: This study was undertaken to determine species distribution, biofilm formation and in-vitro antifungal susceptibility of candida isolated in our tertiary care hospital. METHOD: Eighty seven clinical isolates obtained from urine specimens were subjected to wet mount, Gram's stain and cultured on Sabouraud's Dextrose agar (SDA) medium. Conventional method for yeast identification was done. Biofilm forming ability of each isolate was detected using microtitre plate method. Antifungal susceptibility against posaconazole, amphotericin-B, fluconazole, itraconazole, ketoconazole, 5-flucytosine, voriconazole, and caspofungin was tested using Sensititre® Yeastone® (Trek diagnostic systems). RESULTS AND DISCUSSION: Out of 87 candida isolates, 31.03% (n=27) were C. albicans and 68.97% (n=60) were non albicans candida species (NAC). Among 60 NAC, C. kruseii 29.89% (n=26), C. glabrata 24.14% (n=21), C. tropicalis 14.94% (n=13). Among all isolates, 36.78% (n=32) were biofilm producers and biofilm positivity more among C. albicans 55.56% (n=15) as compared to NAC 28.33% (n=17) (Pvalue<0.002). The maximum positivity was observed with isolates from plastic devices (61.8%). The minimum inhibitory concentrations of all antifungal drugs against all isolates were within susceptible range except for fluconazole which was resistant to C. kruseii. CONCLUSION: C. albicans remains the major isolate from urine samples and also biofilm formation as a virulence factor might have a higher significance for C. albicans than for NAC and its ability to form biofilm is intricately linked with ability of organisms to adhere, colonize and subsequently cause infection.

Ventilator-associated pneumonia in a tertiary care intensive care unit: Analysis of incidence, risk factors and mortality.

BACKGROUND: Ventilator-associated pneumonia (VAP) is the most common nosocomial infection diagnosed in the intensive care unit (ICU) and in spite of advances in diagnostic techniques and management it remains a common cause of hospital morbidity and mortality. OBJECTIVE: The primary objective of the following study is to determine the incidence, various risk factors and attributable mortality associated with VAP and secondary objective is to identify the various bacterial pathogens causing VAP in the ICU. MATERIALS AND METHODS: This prospective observational study was carried out over a period of 1 year. VAP was diagnosed using the clinical pulmonary infection score. Endotracheal aspirate (ETA) and bronchoalveolar lavage (BAL) samples of suspected cases of VAP were collected from ICU patients and processed as per standard protocols. STATISTICAL ANALYSIS: Fisher's exact test was applied when to compare two or more set of variables were compared. RESULTS: The incidence of VAP in our study was 57.14% and the incidence density of VAP was 31.7/1000 ventilator days. Trauma was the commonest underlying condition associated with VAP. The incidence of VAP increased as the duration of mechanical ventilation increased and there was a total agreement in bacteriology between semi-quantitative ETAs and BALs in our study. The overall mortality associated with VAP was observed to be 48.33%. CONCLUSIONS: The incidence of VAP was 57.14%. Study showed that the incidence of VAP is directly proportional to the duration of mechanical ventilation. The most common pathogens causing VAP were Acinetobacter spp. and Pseudomonas aeruginosa and were associated with a high fatality rate.

Prevalence of Pseudomonas aeruginosa in post-operative wound infection in a referral hospital in Haryana, India.

BACKGROUND: The objective of our study was to determine the prevalence of Pseudomonas aeruginosa in the isolates of postoperative wound and its susceptibility pattern to commonly used antibiotics. MATERIALS AND METHODS: During a 2-year period, specimens were received as postoperative wound swabs in Microbiology Laboratory, Maharaja Agrasen Medical College, Agroha (Hisar), Haryana, India. RESULT: Of the 300 bacterial isolates, 89 (29.6%) were P. aeruginosa, followed by Escherichia coli (61, 20.3%), Klebsiella spp. (50, 16.6%), Staphylococcus aureus (43, 14.3%), Proteus spp. (19, 6.3%), Acinetobacter spp. (9, 3.0%), and Citrobacter freundii (2, 0.6%). There was no growth in 27 (9.0%) specimens. CONCLUSION: P. aeruginosa isolation was higher in male patients and most common in the age group of 21-40 years. The susceptibility pattern showed the organism to be most commonly susceptible to imipenem, followed by meropenem, cefoperazone/sulbactam, ticarcillin/clavulanate, and amikacin.

An approach to uropathogenic Escherichia coli in urinary tract infections.

PURPOSE: To study the occurrence and characterization of Uropathogenic Escherichia Coli (UPEC) in cases with urinary tract infections. MATERIALS AND METHODS: A total of 220 symptomatic cases from urinary tract infections and 50 stool samples from apparently healthy individuals were included. The colonies identified as Escherichia Coli were screened for virulence factors, that is, hemolysin, Mannose Resistant and Mannose Sensitive Hemagglutination (MRHA, MSHA), Cell surface hydrophobicity, and Serum resistance. RESULTS: Among the 220 cases 91 (41.36%) were hemolytic, 68 (30.90%) showed MRHA, 58 (26.36%) were cell surface hydrophobicity positive, and 72 (32.72%) were serum-resistant. In 50 controls, three (6%) were hemolytic, six (12%) showed MRHA, nine (18%) showed cell surface hydrophobicity, and 12 (24%) were serum-resistant. The difference between cases and controls for hemolysis and MRHA were significant (P<0.001 and P<0.01, respectively). A total of 14 atypical E. coli were isolated from the urine and all showed the presence of one or the other virulence markers. Out of the 18 mucoid E.coli isolated, 10 were serum-resistant. CONCLUSIONS: The present study revealed that out of 220 urinary isolates, 151 could be labeled as UPEC.