Direct and early detection of Mycobacterium tuberculosis complex and rifampicin resistance from sputum smears.

SETTING: Four district DOTS centres in Sikkim, India. OBJECTIVE: Direct and early detection of Mycobacterium tuberculosis complex and rifampicin (RMP) susceptibility using the INNO-LiPA Rif.TB assay. DESIGN: DNA was extracted from 44 inactivated acid-fast bacilli-positive sputum slides using the Gen Elute bacterial genomic DNA kit. Eluted DNA were ethanol precipitated. The LiPA kit detected RMP susceptibility by detection of mutation in the 81 base pair (bp) rpoB gene. Culture of sputum from the same person was used for drug susceptibility testing for RMP using the proportion method. RESULTS: Amplification of 88.6% (39 slides) was achieved from DNA extracted using kits (66.7% for 1+, 81.8% for 2+ and 100% for 3+ slides). Hybridisation using the LiPA kit was applied to 31 amplified products. There was a concordance of 96.8% (24 RMP-resistant and 6 RMP-susceptible samples) and discordance of 3.2% (one sample resistant using the proportion method and susceptible on LiPA) between LiPA and the proportion method. CONCLUSION: LiPA proved useful on DNA extracted from smear-positive slides. Appropriate treatment regimen could be decided early. Slide smear preparation could replace sputum transport in cetylpyridinium chloride, reducing biohazard and thereby controlling the transmission of multidrug-resistant tuberculosis in the community.

Clinico-etiologic correlates of onychomycosis in Sikkim.

AIMS AND OBJECTIVES: The etiological spectrum of any superficial mycosis is largely dependent on the flora in the immediate environment of the individual. It is influenced by the geographic, climatic and occupational factors. The study is basically to formulate baseline data for the species prevalence of various dermatophytes in patients with onychomycosis attending the Central Referral Hospital, Gangtok, Sikkim. MATERIALS AND METHODS: Thirty-four clinically suspected cases of onychomycosis were subjected to mycological studies. RESULTS: Thirty-two (94.12%) cases were positive for fungal elements by direct microscopy and 28 (82.35%) by culture. Young adults in the age group of 21-30 years were mainly affected. The male:female ratio was 1.125:1. Dermatophytes were isolated in 18 cases (64.29%). Trichophyton tonsurans (44.44%) was the most common isolate followed by T. mentagrophytes (22.22%), T. rubrum (11.11%), T. verrucosum (11.11%) and Microsporum audouinii (11.11%). Apart from dermatophytes, Aspergillus niger (21.43%) and Penicillium marneffei (14.28%) were also isolated. CONCLUSION: Dermatophytes, mainly T. tonsurans, as well as moulds other than dermatophytes were isolated from onychomycosis patients in Gangtok, Sikkim.

Occurrence of pathogenic fungi in soil of burrows of rats and of other sites in bamboo plantations in India and Nepal.

This study examined 215 samples of soil from burrows of rats, other sites in bamboo plantations in different parts of India and Nepal by dilution plating and mouse passage technique for occurrence of Penicillium marneffei and other pathogenic fungi. None of the samples including 25 collected from the burrows of a bamboo rat (Cannomys badius) known to be a carrier of P. marneffei, was positive for the fungus. Among the pathogenic fungi recovered were four isolates of Pseudallescheria boydii (including one from Nepal), two of Trichosporon asteroides, one of Scytalidium hyalinum, 23 isolates of Trichophyton mentagrophytes var. mentagrophytes (including two from Nepal), and two of Microsporum gypseum. Fourteen of the 23 isolates of T. mentagrophytes var. mentagrophytes when tested with the mating types of Arthroderma vanbreuseghemii were found to be of the '+' mating type. The frequent recovery of this dermatophyte from soils of bamboo plantations in several parts of India is remarkable. The study also demonstrates for the first time the occurrence of P. boydii and T. mentagrophytes var. mentagrophytes in Nepalese soil. Among the other fungi recovered were several isolates of species of Aspergillus, Penicillium, Paecilomyces, Fusarium, Chrysosporium, Acremonium, Rhizopus, Mucor, Geotrichum, Trichosporon and Rhodotorula.