Detection of NDM-1 and VIM Genes in Carbapenem-Resistant Klebsiella pneumoniae Isolates from a Tertiary Health-Care Center in Kathmandu, Nepal.

BACKGROUND: Klebsiella pneumoniae is one of the leading causes of nosocomial infections. Carbapenems are used as the last resort for the treatment of multidrug resistant Gram-negative bacterial infections. In recent years, resistance to these lifesaving drugs has been increasingly reported due to the production of carbapenemase. The main objective of this study was to detect the carbapenem-resistant genes blaNDM-1 and blaVIM in K. pneumoniae isolated from different clinical specimens. METHODS: A total of 585 clinical specimens (urine, pus, sputum, blood, catheter tips, and others) from human subjects attended at Annapurna Neurological Institute and Allied Sciences, Kathmandu were obtained in the period between July 2018 and January 2019. The specimens were isolated and identified for K. pneumoniae. All K. pneumoniae isolates were processed for antimicrobial susceptibility testing (AST) using the disk diffusion method. The isolates were further phenotypically confirmed for carbapenemase production by the modified Hodge test (MHT) using imipenem (10 µg) and meropenem (10 µg) discs. Thus, confirmed carbapenemase-producing isolates were further screened for the production of blaNDM-1 and blaVIM using conventional polymerase chain reaction (PCR). RESULTS: Among the clinical isolates tested, culture positivity was 38.29% (224/585), and the prevalence of K. pneumoniae was 25.89% (58/224). On AST, K. pneumoniae exhibited resistance toward carbapenems including ertapenem, meropenem, and imipenem, while it showed the highest susceptibility rate against to tigecycline (93.1%; 54/58). Overall, AST detected 60.34% (35/58) carbapenem-resistant isolates, while the MHT phenotypically confirmed 51.72% (30/58) isolates as carbapenemase-producers and 48.28% (28/58) as carbapenemase nonproducers. On subsequent screening for resistant genes among carbapenemase-producers by PCR assay, 80% (24/30) and 3.33% (1/30) isolates were found to be positive for blaNDM-1 and blaVIM, respectively. In the same assay among 28 carbapenem nonproducing isolates, 9 (32.14%) isolates were positive for blaNDM-1 gene while none of them were tested positive for blaVIM gene. CONCLUSIONS: Molecular detection of resistant genes provides greater specificity and sensitivity than those with conventional techniques, thus aiding in accurate identification of antimicrobial resistance and clinical management of the disease.

Prevalence of Colistin-resistant Gram-negative Isolates Carrying the mcr-1 Gene among Patients Visiting a Tertiary Care Center.

INTRODUCTION: Gram-negative isolates harboring mobilized colistin resistance (mcr-1) gene are a great threat to human health. They have been reported worldwide among various bacterial isolates. This work aimed to study the prevalence of colistin resistance among Gram-negative bacteria and the incidence of mcr-1 gene among these isolates. METHODS: A descriptive cross-sectional study was done at a tertiary care center from June 2016 to February 2017. An ethical approval was taken from review board of the Nepal Health Research Council (Reg. no: 274/2016). Convenience sampling was used. The data was collected and analyzed using Microsoft Excel 2010 and Statistical Package for Social Sciences (SPSS) Version 16 . Point estimate at 95% Confidence Interval was calculated along with frequency and proportion for binary data. RESULTS: Among 485 gram-negative isolates, only 13 (2.68%) (1.26-6.62 at 95% Confidence Interval) isolates were colistin-resistant and mcr-1 was present in two isolates. Predominant colistin-resistant isolates were E. coli 6 (4.1%), Enterobacter spp 2 (2.81%), and Acinetobacter spp 2 (2.81%). A high level of colistin-resistance was noted in 4 (30.7%) isolates as indicated by the very high value of colistin MIC (>256 µg/ml). ICU was the major site of isolation of colistin-resistant and mcr-1 positive pathogens. The majority of colistin-resistant isolates were highly drug-resistant and were sensitive only to polymyxin B. Antibiotics like imipenem, amikacin, gentamicin, aztreonam, ciprofloxacin, and piperacillin-tazobactam were effective for few of these isolates. CONCLUSIONS: Though the prevalence of mcr-1 gene was low among colistin-resistant gram-negative isolates, the resistant pattern was quite alarming as these isolates were highly drug-resistant.

Evaluation of drug promotional materials in a hospital setting in Nepal.

OBJECTIVES: Unethical drug promotion is a common problem worldwide. In, Nepal, there is limited vigilance on the quality of information supplied by the drug companies to the doctors. The objectives of this study were to analyze the promotional materials provided by the drug companies as per WHO ethical criteria for medicinal drug promotion. METHODS: Promotional materials present in the Drug Information Center (DIC) during the period from September to December 2007 were collected. The collected promotional materials of different pharmaceutical companies were compared with WHO's Ethical Criteria for Medicinal Drug Promotion. RESULTS: The name of active constituent(s) was mentioned in 87.87% (n=29) of promotional items. Therapeutic indication was mentioned in 87.88% (n=29) of promotional material but information on side effects [33.33% (n=11)], drug interactions [9.09% (n=3)] and use in pregnancy and lactation [12.12% (n=4)] were lacking in the majority of promotional materials. CONCLUSION: In a country like Nepal with limited drug information resources, the promotional materials provided by the manufacturers can largely influence the prescription behavior of the clinicians. Our findings suggest the need for interventions to improve the content of the promotional materials provided by the drug companies.