Performance evaluation of the new Roche cobas AmpliPrep/cobas TaqMan HCV test, version 2.0, for detection and quantification of hepatitis C virus RNA.

To evaluate the analytical performance and explore the clinical applicability of the new Roche cobas AmpliPrep/cobas TaqMan HCV test, v2.0 (CAP/CTM v2.0), a platform comparison was performed on panels and diagnostic samples with the Roche cobas AmpliPrep/cobas TaqMan HCV test (CAP/CTM v1.0), the Siemens Versant HCV RNA 3.0 branched DNA (bDNA) test, the Abbott m2000 RealTime HCV assay (Realtime assay), and the Siemens Versant HCV transcription-mediated amplification (TMA) test (TMA assay). The analytical performance of the CAP/CTM v2.0 on WHO and Acrometrix panels and clinical specimens of patients infected with HCV genotype 1, 2, 3, 4, 5, or 6 relative to that of the CAP/CTM v1.0 was significantly improved. In a qualitative comparison of the CAP/CTM v2.0 relative to the TMA assay on genotype 1 to 4 samples, the two tests proved to be almost equally sensitive. Response-guided therapy in one of five HCV genotype 4-infected patients previously tested with the CAP/CTM v1.0 would have significantly changed if tested with the CAP/CTM v2.0. In conclusion, the Roche CAP/CTM v2.0 has significantly better performance characteristics than the former CAP/CTM HCV v1.0 and the bDNA assay and performance characteristics comparable to those of the Realtime assay.

Evaluation of hospital environment for presence of Mucorales during COVID-19-associated mucormycosis outbreak in India - a multi-centre study.

BACKGROUND: An unprecedented rise in the number of COVID-19-associated mucormycosis (CAM) cases has been reported in India. Myriad hypotheses are proposed for the outbreak. We recently reported uncontrolled diabetes and inappropriate steroid therapy as significant risk factors for the outbreak. However, Mucorales contamination of hospital environment was not studied. AIM: To perform a multi-centre study across India to determine possible Mucorales contamination of hospital environment during the outbreak. METHODS: Eleven hospitals from four zones of India representing high to low incidence for mucormycosis cases were included in the study. Samples from a variety of equipment used by the patients and ambient air were collected during May 19th, 2021 through August 25th, 2021. FINDINGS: None of the hospital equipment sampled was contaminated with Mucorales. However, Mucorales were isolated from 11.1% air-conditioning vents and 1.7% of patients' used masks. Other fungi were isolated from 18% of hospital equipment and surfaces, and 8.1% of used masks. Mucorales grew from 21.7% indoor and 53.8% outdoor air samples. Spore counts of Mucorales in air were significantly higher in the hospitals of North and South zones compared to West and East zones (P < 0.0001). Among Mucorales isolated from the environment, Rhizopus spp. were the most frequent genus. CONCLUSION: Contamination of air-conditioning vents and hospital air by Mucorales was found. Presence of Mucorales in these areas demands regular surveillance and improvement of hospital environment, as contamination may contribute to healthcare-associated mucormycosis outbreaks, especially among immunocompromised patients.

Potential of mycobacterial excretory secretory protein antigens (sevatb ES-31, ES-43, EST-6 and ES-20) as biomarkers to detect Mycobacterium tuberculosis bacilli.

There is a need for a simple and reliable method to identify Mycobacterium tuberculosis from nontuberculous mycobacteria (NTM). The utility of mycobacterial ES-31, ES-43, EST-6 or ES-20 antigen as a biomarker for differentiation of Mycobacterium tuberculosis bacilli from nontuberculous mycobacteria was explored using Fluorescein isothiocyanate conjugated antibodies against these antigens. Detection of these antigens was done from M.tb H(37)Ra and H(37)Rv DSS antigen. The presence of antigen in bacilli using FITC labelled antibody was indicated by green fluorescence on the cell surface while, its absence by no fluorescence under microscope. In M.tb H(37)Ra and H(37)Rv bacilli, fluorescence was observed on addition of FITC labelled anti ES-31 and anti ES-43 antibody; whereas no fluorescence was observed in case of EST-6 and ES-20 antibody conjugates. However all the antigens were detected in detergent soluble sonicate antigen of tubercle bacilli on addition of FITC conjugates. Fluorescence was not observed for ES-31, ES-43, EST-6 and ES-20 antigen in any of the tested NTM as well as in Escherichia coli. SEVA TB ES-31 and ES-43 may be used as biomarkers to distinguish M.tuberculosis bacilli from NTM.

Mycotic keratitis due to Engyodontium album: first case report from India.

Engyodontium album is a rare and an unusual human pathogen. It is a common inhabitant of waste and moist material and frequently isolated from substrates such as paper, jute, linen and painted walls. This fungus grew within 3 days on SDA with chloramphenicol from corneal scrapping of a 70-year-old male farmer with a history of trauma by unknown vegetative matter. The fungus can be confused with Tritirachium sp and Beauveria sp.

Mycobacterial ES-31 serine protease--a biomarker for mycobacterium tuberculosis--a preliminary report.

There is a need for simple and reliable method to identify Mycobacterium tuberculosis from AFB smear positive cases. Utility of mycobacterial ES-31 serine protease as a marker to detect Mycobacterium tuberculosis bacilli was explored using Fluorescein isothiocyanate conjugated anti-ES-31 serine protease antibody. The presence of ES-31 serine protease in bacilli was indicated by green fluorescence on the cell surface. Green fluorescence was observed with M.tb.H37Ra bacilli and M.tb.H37Rv bacilli while no Fluorescence was observed with M. chelonae, Nocardia farcinicum as well as in E. coli showing the usefulness of ES-31 serine protease as a marker for identification of mycobacterium tubercle bacilli in cultures.

Status of high level aminoglycoside resistant Enterococcus faecium and Enterococcus faecalis in a rural hospital of central India.

Considering the emergence of high level aminoglycoside resistance (HLAR) in enterococci this study was undertaken to determine their status in a rural setting. HLAR by disc diffusion and agar dilution, beta lactamase by nitrocefin disc and vancomycin resistance by agar dilution was determined in 150 enterococcal isolates, as per NCCLS guidelines. Only two species, Enterococcus faecalis (85.5%) and Enterococcus faecium (14.7%) were recovered, mostly from blood. Forty six percent showed HLAR. Multi drug resistance and concomitant resistance of HLAR strains to beta lactams were quite high. None showed beta lactamase activity or vancomycin resistance.

Candida colonization in preterm babies admitted to neonatal intensive care unit in the rural setting.

PURPOSE: Candida colonization in neonates results in significant morbidity and mortality. The purpose of this study was to determine colonization of Candida spp. in preterm babies and identify the risk factors. METHODS: Swabs from oral, rectum, groin and umblicus of 103 preterm and 100 term neonates were obtained within 24 hours of birth, day three, day five, day seven and thereafter every week till the neonate was admitted in the neonatal intensive care unit (NICU). Swabs were also collected from the mother's vagina prior to delivery. Twice every month, air of the NICU was sampled by settle plate and swabs were collected from the hands of health care workers and inanimate objects of NICU. Identification and speciation was done by standard methods. Antibiotic sensitivity was studied against amphotericin B, ketoconazole and fluconazole by disk diffusion method. RESULTS: Colonization with Candida was significantly higher in preterms. Earliest colonization was of oral mucosa and 77.1% of the preterms had colonised at various sites by the first week of life. Significant risk factors in colonized versus non-colonized preterms were male sex, longer duration of rupture of membranes (DROM), administration of steroids and antibiotics and vaginal colonization of mothers, whereas those in preterms versus terms were low birth weight and gestational age. C. albicans was the commonest species, both in the colonized preterms (45.9%) and vagina of mothers. Resistance was seen to fluconazole and ketoconazole only. No Candida spp. was isolated from health care personnel or environment. CONCLUSIONS: Colonization of preterms by Candida is a significant problem in NICU and the significant risk factors observed in colonized preterms were male sex, longer DROM, administration of steroids and antibiotics and vaginal colonization of mothers.

Keratitis due to Colletotrichum dematium--a case report.

Colletotrichum dematium has been rarely reported from India before. The present case, a farmer, developed peripheral corneal ulcer five days following trauma with plant. At presentation his visual acuity was 6/60 (unaided) and 6/24P with pinhole. Slit lamp and fluorescent stain examination revealed paracentral corneal ulcer with irregular margins, stromal infiltration and multiple epithelial defects. Microbiological examination of corneal samples confirmed the initial diagnosis of fungal corneal ulcer and the fungus was identified as C.dematium. Patient was treated with topical natamycin and ciprofloxacin. Patient left against medical advice and was lost to follow up. This report emphasizes that Colletotrichum keratitis may not be rare. Early diagnosis may help in institution of specific therapy early in the disease.