Contaminated antiseptics--an unnecessary hospital hazard

A total of 284 antiseptic solutions were studied to check for their sterility. The overall antiseptic contamination rate was 15.14%. 14.85% of freshly prepared antiseptics were contaminated. Here, the problem could be attributed to inadequate precautions while preparing the antiseptics. 15.3% of the in-use antiseptics were contaminated. This could be due to improper handling. Non-fermenters (45.45%), Pseudomonas aeruginosa (30.30%) and Klebsiella spp. (22.72%) were the commonest organisms recovered from the antiseptics. In 44.44% of patients, the isolates obtained from the catheterised urine in the same wards matched with the isolates from antiseptics of that ward. Antiseptic solutions have to be regularly monitored. If they are found to be contaminated, they should be discarded immediately and replaced by fresh sterile antiseptics otherwise instead of preventing infection, antiseptics will become a source of hospital-acquired infection.     

Microdissection genotyping of archival fixative treated tissue for Gaucher disease

The genetic diagnosis of Gaucher disease by molecular methods is complicated by the existence of a highly homologous transcribed pseudogene (96% identity) that is found in close proximity to the true gene on chromosome 1q21. In addition, the pseudogene sequence can mimic disease-causing mutations in the true gene. Selective polymerase chain reaction (PCR) amplification of the true gene can be accomplished in extracted DNA from fresh-frozen samples by designing oligonucleotide primers to hybridize to defined regions that are not present in the pseudogene. This standard molecular approach, which entails amplification of relatively long segments of intact DNA, is not feasible in archival, paraffin-embedded, solid-tissue specimens in which the negative effects of chemical fixation result in DNA strand scission and breakdown of nucleic acid. A novel approach, specifically created for use with archival, fixative-treated tissue specimens, was developed for detection and characterization of common mutations of Gaucher disease. Three separate robust PCR reactions were formulated, 2 for selective amplification of portions of only the true gene exons 2 and 9, with a third reaction targeting exon 10, wherein both the true and pseudogene were coamplified. In the latter, DNA sequencing was used to determine the presence of true and pseudogene allele content in addition to identification of base sequence alterations. This method, requiring a single, 4-microm-thick histologic section, was successfully applied to archival paraffin block tissue specimens that had been in storage for up to 75 years. It was capable of accurately genotyping common Gaucher disease mutations as well as discovering a novel mutation and genetic polymorphism. We recommend our approach when only fixative-treated tis sue is available for molecular genotyping.     

Epidemiology of chronic suppurative otitis media and deafness in a rural area and developing an intervention strategy

Of 613 children evaluated in a village in Haryana 94 (15.3%) were observed to have chronic suppurative otitis media (CSOM). Fifty eight (61.7%) children had hearing impairment. CSOM contributed to 71.6% of the hearing impaired (58/81). On analysis of association of CSOM with literacy and socio-economic status of mothers, and age, sex, and upper respiratory tract infections (URI) in children positive correlation was observed only with URIs (P<0.001). Literacy and socio-economic status of the mothers did not correlate significantly with knowledge about treatment seeking, and ear cleaning practices, probably due to the narrow range of incomes and literacy levels. An intervention program consisting of play, demonstrations, health charts and slogans, and aural cleaning and antibiotic drops was introduced.     

Intramuscular inoculation of Sin Nombre hantavirus cDNAs induces cellular and humoral immune responses in BALB/c mice.

To examine whether genetic immunization with Sin Nombre (SN) hantavirus genes could elicit immune responses, nine fragments spanning the envelope glycoprotein genes G1 and G2, and the complete N gene were cloned into a CMV expression vector. To ensure representation of all potential epitopes, adjacent fragments of the glycoprotein genes overlapped one another by 100 nucleotides. Vectors containing the gene fragments were inoculated intramuscularly into BALB/c mice and splenocyte proliferation and western blot-detectable antibodies and neutralization titers were determined. The N gene and seven of the nine M segment-derived cDNAs tested produced significant specific lymphoproliferative responses, and many of the constructs elicited either neutralizing or western blot-detectable antibodies. These promising results encourage the development of infection models for SN virus that will be capable of detecting protective responses.     

Shigellosis in Bay of Bengal Islands,India: clinical and seasonal patterns,surveillance of antibiotic susceptibility patterns,and molecular characterization of multidrug-resistant Shigella strains isolated during a 6-year period from 2006 to 2011

This study aims to determine the clinical features and seasonal patterns associated with shigellosis, the antimicrobial resistance frequencies of the isolates obtained during the period 2006–2012 for 22 antibiotics, and the molecular characterization of multidrug-resistant strains isolated from endemic cases of shigellosis in the remote islands of India, with special reference to fluoroquinolone and third-generation cephalosporins resistance. During the period from January 2006 to December 2011, stool samples were obtained and processed to isolate Shigella spp. The isolates were evaluated with respect to their antibiotic resistance pattern and various multidrug resistance determinants, including resistance genes, quinolone resistance determinants, and extended-spectrum β-lactamase (ESBL) production. Morbidity for shigellosis was found to be 9.3 % among children in these islands. Cases of shigellosis occurred mainly during the rainy seasons and were found to be higher in the age group 2–5 years. A wide spectrum of resistance was observed among the Shigella strains, and more than 50 % of the isolates were multidrug-resistant. The development of multidrug-resistant strains was found to be associated with various drug-resistant genes, multiple mutations in the quinolone resistance-determining region (QRDR), and the presence of plasmid-mediated quinolone-resistant determinants and efflux pump mediators. This report represents the first presentation of the results of long-term surveillance and molecular characterization concerning antimicrobial resistances in clinical Shigella strains in these islands. Information gathered as part of the investigations will be instrumental in identifying emerging antimicrobial resistance, for developing treatment guidelines appropriate for that community, and to provide baseline data with which to compare outbreak strains in the future.