Browsing by Author "Khanal, Laxmi Kant"
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Publication Phenotypic and Genotypic Detection of Metallo-b Lactamase in Non-Fermenting Gram Negative Bacilli Obtained from Clinical Samples in a Tertiary Care Hospital in Nepal(Institute of Medicine, Tribhuvan University, 2025) Sapkota, Jyotshna; GC, Divya Shree; Adhikari, Ram Prasad; Khadka, Shusila; Pandey, Ritu; Sah, Anil Kumar; Khanal, Laxmi KantAbstract: Introduction Non-fermenting Gram-negative bacilli, including Pseudomonas aeruginosa and Acinetobacter baumannii, are significant nosocomial pathogens with limited treatment options due to their intrinsic and acquired resistance mechanisms. Among these, metallo-β-lactamases are of major concern which hydrolyze carbapenems and contribute to antimicrobial resistance. This study aimed to determine the prevalence of Metallo-β-Lactamase-producing non-fermenting gram negative bacilli in clinical specimens using both phenotypic and genotypic methods. Methods A descriptive cross-sectional study was conducted at Nepal Medical College Teaching Hospital from January 2024 to December 2024. A total of 16,954 clinical specimens were processed for culture and sensitivity testing. NFGNB were identified using standard microbiological methods. Antimicrobial susceptibility testing was conducted using the Kirby-Bauer disk diffusion method following Clinical and Laboratory Standards Institute guidelines. MBL production was detected phenotypically using the Imipenem-EDTA combined disc method. The presence of IMP and VIM genes was confirmed by conventional PCR. Results Among 16,954 specimens, 163 (0.96%) NFGNB isolates were identified, with Pseudomonas aeruginosa (52.1%) being the most prevalent, followed by Acinetobacter species (39.3%) and Burkholderia species (8.6%). MBL production was detected in 22 (13.5%) isolates: Pseudomonas aeruginosa (59.1%), and Acinetobacter species (40.9%). Genotypically IMP and VIM genes were found in 36.4% and 31.8% of MBL-positive isolates, respectively, while one isolate harbored both genes. Notably, 27.3% of phenotypic MBL producers tested negative for both IMP and VIM, suggesting the potential involvement of other MBL genes. Conclusion The significant prevalence of MBL-producing NLFGNB, particularly among Pseudomonas aeruginosa and Acinetobacter species, highlights a serious challenge for antimicrobial therapy and underscores an urgent need for robust infection control and antimicrobial stewardship strategies.Publication Seroprevelance of Dengue Among Healthy Blood Donors in Blood bank(Nepal Health Research Council, 2024) Sapkota, Jyotshna; Thapa, Ranjan; Nepal, Basudha; Adhikari, Ram Prasad; Khadka, Sushila; Aryal, Kiran; Acharya, Avinav; Kunwar, Saroj; Khanal, Laxmi KantBackground: Dengue is a viral infection, commonly found in tropical and subtropical regions across the globe particularly in urban and semi-urban settings. There has been significant increase in dengue cases over past two decades. This study aims to determine the prevalence of dengue NS1 antigen, IgM, and IgG antibodies among healthy blood donors at the Nepal Medical College Blood Bank. Methods: The descriptive cross-sectional study was carried out at the blood bank of Nepal Medical College between August to November 2023. Ethical approval was received from the Institutional Review Committee of Nepal Medical College (Ref: -080/081). Two hundred and thirty two serum samples from the healthy blood donors were screened with Dengue RDT (SD Bioline Dengue Duo) for detection of NS1 antigen, IgM and Ig G antibodies. Results: Among the samples 18/232 (7.75%) were found IgM positive and 25/232 (10.77%) were found IgG positive. Similarly, 12/232 (5.17%) showed both IgG and IgM positive; however, all sample tested negative for NS1 Conclusions: Detection of Dengue antibodies in asymptomatic blood donors in endemic areas indicates a potential risk for recipients to contract dengue infection through blood transfusion. Implementing a screening program during dengue outbreaks could effectively mitigate this risk. Keywords: Blood donors; dengue; seroprevalence.Publication Speciation and Fluconazole Susceptibility of Candida Isolates from Clinical Samples in a Tertiary Hospital in Nepal(Institute of Medicine, Tribhuvan University, 2024) Sapkota, Jyotshna; Aryal, Kiran; Adhikari, Ram Prasad; Khadka, Sushila; Pandey, Ritu; Khanal, Laxmi Kant; Shrestha, Deepti; Sharma, ManishaAbstract: Introduction Candida species, commonly present as human commensals, are notable opportunistic pathogens responsible for superficial and invasive infections, particularly in hospitalized patients. This study aimed to characterize the distribution and antifungal susceptibility patterns of Candida species isolated from various clinical specimens at a tertiary care hospital in Nepal. Methods A descriptive cross-sectional, laboratory-based study was conducted in the Clinical Microbiology Laboratory of Nepal Medical College Teaching Hospital between July 2023 and June 2024. Clinical specimens yielding Candida species over the one-year period were included. Growth on Sabouraud dextrose agar was analyzed for colony morphology, Gram staining, germ tube production, and urea hydrolysis. Speciation of Candida isolates was performed using CHROMagar. Additionally, fluconazole susceptibility testing was conducted following Clinical and Laboratory Standards Institute guidelines. Results A total of 72 Candida isolates were identified from samples such as sputum, urine, and high vaginal swabs. Candida albicans was the most prevalent species 45 (62.5%), followed by non-albicans Candida (NAC) species, including C. glabrata 14 (19.4%), 9 C. tropicalis (12.5%), and 4 C. parapsilosis (5.6%). Fluconazole resistance was observed in 22.2% of total isolates of Candida, with C. glabrata exhibiting the highest resistance 12, (86%), whereas 4 (9%) of C. albicans demonstrated resistance to it. Conclusion The findings highlight a shift in epidemiology toward NAC species and raise concerns about rising fluconazole resistance. These results underscore the need for routine species identification and antifungal susceptibility testing to guide effective management strategies. The study advocates for antifungal stewardship programs and ongoing surveillance to address emerging resistance trends in Candida infections.