Aim: The extent of methicillin-resistant Staphylococcus aureus (MRSA) infection in Nepalese children is largely unknown. Materials & methods: Six hundred and seventy-two clinical samples collected from 232 patients between June and November 2016 were processed in a microbiology laboratory. Results: Out of 300 culture-positive samples, 52 (17.3%) were S. aureus isolates. Among those 52, 39 (75.0%) were found to be MRSA. The infection rate of S. aureus was shown to be higher in inpatients (55.7%) compared with outpatients (44.3%) at p = 0.637, 95% CI. Thirteen types of antibiotics were used in the antibiotic susceptibility test. MRSA isolates showed 100 and 0% resistance to penicillin and vancomycin, respectively. The D-test showed inducible clindamycin-resistant phenotype in 15.4% of MRSA isolates. Conclusion: This demonstrates the utmost need for routine testing for MRSA in Nepalese hospitals.

Background: Staphylococcus aureus is one of the important superbugs distributed throughout the world. It causes minor skin infections to severe complications including nosocomial infections in both hospitals and community settings. These strains have multi-drug resistant property. Hence, they are difficult to manage which increase health-related costs and simultaneously intensifying the need for new antibiotics. The extent of Methicillin-Resistant Staphylococcus aureus (MRSA) in children is largely unknown. The study determines the current status of S. aureus and MRSA causing various infections in pediatric patients visiting International Friendship Children’s Hospital (IFCH). Methods: A cross-sectional study was conducted among patients visiting a hospital. Various clinical specimens were aseptically collected and processed according to standard microbiological procedures. Isolation and identification of S. aureus were done by microscopy, mannitol fermentation, and coagulase positivity. All identified S. aureus isolates subjected to in-vitro antibiogram by Kirby-Bauer disc diffusion technique adopting Clinical and Laboratory Standards Institute (CLSI) guideline. Isolates resistant to cefoxitin were considered to be MRSA. Whereas, isolates produced D-shaped inhibition zone around clindamycin when kept near erythromycin were considered to be Inducible Clindamycin Resistant (ICR). Results: 672 various types of clinical samples were processed from the microbiology laboratory from June and November 2015. Out of 300 culture positive samples, 52 (17.3%) were S. aureus isolates, among them 39 (75.0%) were found to be MRSA. The D-test showed that Macrolide-Lincosamide-Streptogramin-B (MLSB) phenotype was 15.4%. Conclusion: The study shows the MRSA occurrence is prevalent in pediatric patients and newer classes’ drugs are found more effective than β-lactam drugs to treat S. aureus infection. However, restriction on the indiscriminate use of such drugs may be an effective strategy to control the drug resistance. Keywords: Methicillin-Resistant Staphylococcus aureus (MRSA), Macrolide-Lincosamide-Streptogramin B (MLSB) phenotype, Inducible Clindamycin Resistant (ICR) test or D-zone test, Antibiotic resistance, Nepal

Freeze drying (Lyophilization) performed at temperature and pressure below the triple point is being practiced for the preservation of virus stocks for longer periods. The present study is aimed to lyophilize adenovirus strain to study its effects on infectivity and viral load. In-house adenovirus reference strain (stock virus) was propagated in Hep-2 cell line in 25cm2 cell culture flasks. In 24-well plates the serial dilutions of stock virus from 10-1 to 10-7 (100μl inoculum) was inoculated in each well with Hep-2 cells for TCID50 titer and viral DNA was extracted separately to determine viral load by Taqman Real Time PCR. Stock virus was lyophilized in 3 lots and stored at RT (25±2°C) and 4°C separately for 1, 4 and 6 months and subjected to TCID50 (for viral infectivity) and viral load assay (for total viral genome copies). Following lyophilisation and storage of adenoviral strains at RT and 4°C separately did not affect significantly on the viral stability, infectivity as well as viral copy number till 4 months. However, storage at RT for 6 months resulted in 1 log reduction in viral copy number. Thus, storage of even lyophilized virus stock would necessitate a temperature of at least 4°C for prolonged periods. The present study could successfully lyophilize adenovirus and retain its infectivity over a period of 6 months when stored at RT and 4°C. No significant difference in the infectivity or TCID50 titer was observed in the lyophilized virus as compared to the stock virus. However, the viral load was observed to increase with lyophilization of the virus over 6 months when stored at 4°C which possibly is due to the concentration of the virus on freeze-drying.Nepal Journal of Biotechnology. Dec. 2015 Vol. 3, No. 1: 15-21