Komal Raj Rijal

Multidrug resistant Vibrio cholerae O1 from clinical and environmental samples in Kathmandu city

BackgroundCholera, an infectious disease caused by Vibrio cholerae, is a major public health problem and is a particularly burden in developing countries including Nepal. Although the recent worldwide outbreaks of cholera have been due to V. cholerae El Tor, the classical biotypes are still predominant in Nepal. Serogroup O1 of the V. cholerae classical biotype was the primary cause of a cholera outbreak in Kathmandu in 2012. Thus, this study was designed to know serotypes and biotypes of V. cholerae strains causing recent outbreak with reference to drug resistant patterns. Moreover, we also report the toxigenic strains of V. cholerae from both environmental and clinical specimens by detecting the ctx gene.MethodsTwenty four V. cholerae (n = 22 from stool samples and n = 2 from water samples) isolated in this study were subjected to Serotyping and biotyping following the standard protocols as described previously. All of the isolates were tested for antimicrobial susceptibility patterns using the modified Kirby-Bauer disk diffusion method as recommended by CLSI guidelines. The screening of the ctx genes (ctxA2-B gene) were performed by PCR method using a pair of primers; C2F (5′-AGGTGTAAAATTCCTTGACGA-3′) and C2R (5′-TCCTCAGGGTATCCTTCATC-3′) to identify the toxigenic strains of V. cholerae.ResultsAmong twenty four V. cholerae isolates, 91.7% were clinical and 8.3% were from water samples. Higher rate of V. cholerae infection was found among adults of aged group 20–30 years. All isolates were serogroups O1 of the V. cholerae classical biotype and sub serotype, Ogawa. All isolates were resistant to ampicillin, nalidixic acid and cotrimoxazole. 90.9% were resistant to erythromycin however, tetracycline was found to be the most effective drug for the isolates. All isolates were multidrug resistant (MDR) and possessed a ctx gene of approximately 400 base pairs indicating the toxigenic strains.ConclusionHundred percent strains of V. cholerae were MDR possessing a ctx gene. It suggests that toxigenic strains be identified and proper antibiotic susceptibility testing be conducted. This will allow effective empirical therapy to be used to treat and control cholera.

Re-emergence of susceptibility to conventional first line drugs in Salmonella isolates from enteric fever patients in Nepal

INTRODUCTION Enteric fever is endemic in Nepal and poses a significant public health burden. The first-line drugs ampicillin, chloramphenicol, and cotrimoxazole have not been part of empirical therapy for two decades due to the development of multidrug-resistant Salmonella strains. The objective of this study was to determine the antibiogram pattern of Salmonella serovars isolated from the blood of clinically suspected enteric fever patients. METHODOLOGY A cross sectional study was carried out in a tertiary care hospital in Lalitpur, Nepal, between July 2011 and February 2012. Standard microbiological procedures were followed during collection and processing of blood samples, isolation and identification of Salmonella serotypes. The antimicrobial sensitivity of ampicillin, chloramphenicol, cotrimoxazole, nalidixic acid, and ciprofloxacin was determined using a modified Kirby-Bauer disk diffusion method as per the guidelines of the Clinical and Laboratory Standards Institute. RESULTS Out of 86 Salmonella isolates, 56 (65.1%) were Salmonella Typhi and 30 (34.9%) were Salmonella Paratyphi A. Salmonella Typhi were 100% sensitive to chloramphenicol, cotrimoxazole, and ciprofloxacin and 98.2% sensitive to ampicillin. Similarly, Salmonella Paratyphi A isolates were 100% sensitive to ampicillin and cotrimoxazole and 96.7% sensitive to chloramphenicol and ciprofloxacin. More than 90.0% of isolates were nalidixic acid resistant and none of the Salmonella isolates were multi-drug resistant. CONCLUSIONS This study revealed the increasing frequency of nalidixic acid-resistant Salmonella isolates, indicating the possibility of fluoroquinolone resistance in near future. Furthermore, re-emergence of susceptibility to conventional first-line drugs ampicillin, chloramphenicol, and cotrimoxazole supports the possibility of using these drugs in empirical therapy.

Diagnostic Accuracy of GeneXpert MTB/RIF Assay in Comparison to Conventional Drug Susceptibility Testing Method for the Diagnosis of Multidrug-Resistant Tuberculosis

Xpert MTB/RIF assay is regarded as a great achievement of modern medicine for the rapid diagnosis of multidrug-resistant tuberculosis (MDR-TB). The main purpose of this study was to determine the performance of Xpert MTB/RIF assay compared to conventional drug susceptibility testing (DST) method for the diagnosis of MDR-TB. A comparative cross sectional study was carried out at German-Nepal Tuberculosis Project, Kathmandu, Nepal, from April 2014 to September 2014. A total of 88 culture positive clinical samples (83 pulmonary and 5 extra-pulmonary) received during the study period were analyzed for detection of multidrug-resistant tuberculosis by both GeneXpert MTB/RIF assay and conventional DST method. McNemar chi square test was used to compare the performance of Xpert with that of DST method. A p-value of less than 0.05 was considered as statistically significant. Of total 88 culture positive samples, one was reported as invalid while 2 were found to contain nontuberculous Mycobacteria (NTM). Among remaining 85 Mycobacterium tuberculosis culture positive samples, 69 were found to be MDR-TB positive by both methods. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of GeneXpert MTB/RIF assay were found to be 98.6%, 100%, 100% and 93.8% respectively. Statistically, there was no significant difference between the diagnostic performance of Xpert and conventional DST method for detection of MDR-TB. GeneXpert MTB/RIF assay was found to be highly sensitive, specific and comparable to gold standard conventional DST method for the diagnosis of MDR-TB.

Burden estimation of dengue at National Public Health Laboratory, Kathmandu

Abstract Objective To determine the burden of dengue in the patients visiting National Public Health Laboratory, Kathmandu. Methods A cross sectional study was carried out at National Public Health Laboratory, Kathmandu from May to December, 2013. Serum samples were collected from patients suspected of dengue virus infection and tested by ELISA. Results Among 266 patients suspected of dengue virus infection, 45 (16.9%) showed anti-dengue immunoglobulin M antibodies in serum. Males and economically active people were more infected and the maximum number of cases was during the month of October. Conclusions This study revealed that the proportion of dengue was more in Kathmandu, especially among the economically active males. So, the control measures should be initiated targeting these groups of people.

Epidemiology of Plasmodium vivax Malaria Infection in Nepal

Abstract. Malaria is endemic in the southern plain of Nepal which shares a porous border with India. More than 80% cases of malaria in Nepal are caused by Plasmodium vivax. The main objective of this study was to review the epidemiology of P. vivax malaria infections as recorded by the national malaria control program of Nepal between 1963 and 2016. National malaria data were retrieved from the National Malaria program in the Ministry of Health, Government of Nepal. The epidemiological trends and malariometric indicators were analyzed. Vivax malaria has predominated over falciparum malaria in the past 53 years, with P. vivax malaria comprising 70–95% of the annual malaria infections. In 1985, a malaria epidemic occurred with 42,321 cases (82% P. vivax and 17% Plasmodium falciparum). Nepal had experienced further outbreaks of malaria in 1991 and 2002. Plasmodium falciparum cases increased from 2005 to 2010 but since then declined. Analyzing the overall trend between 2002 (12,786 cases) until 2016 (1,009 cases) shows a case reduction by 92%. The proportion of imported malaria cases has increased from 18% of cases in 2001 to 50% in 2016. The current trends of malariometric indices indicate that Nepal is making a significant progress toward achieving the goal of malaria elimination by 2025. Most of the cases are caused by P. vivax with imported malaria comprising an increasing proportion of cases. The malaria control program in Nepal needs to counter importation of malaria at high risk areas with collaborative cross border malaria control activities.

Status of inducible clindamycin resistance among macrolide resistant Staphylococcus aureus

Clindamycin has long been an option for treating both methicillin sensitive Staphylococcus aureus (MSSA) and methicillin resistant S. aureus (MRSA) infections. So, it is utmost important to perform the susceptibility test for erythromycin and clindamycin. And, there is concern on use of this antibiotic in the presence of erythromycin resistance because of the possibility of induction of cross-resistance among members of macrolide, lincosamide and streptogramin B (MLSB) group. During August 2011 to May 2012, a total of 207 isolates of S. aureus were isolated and among which 29.47% (61) isolates were confirmed as MRSA by cefoxitin (30 µg) disc. All the isolates were further processed for MLSB resistance test by double disc diffusion test of erythromycin (2 µg) and clindamycin (15 µg) at a distance of 15 and 22 mm between them. This study result show 12.56% (26) and 14.49% (30) of inducible macrolide-lincosamide-streptogramin B phenotype (iMLSB) resistance type at 22 and 15 mm disc distance, respectively, showing 15 mm disc distance is potential than 22 mm and 17.39% (36) of cMLSB resistance type. Similarly, both iMLSB and cMLSB are greater in MRSA than MSSA and constitutes 18.05 (11) and 36.06% (22), respectively. Thus, this study concludes that D-test should be used as a mandatory method and is more potential in 15 mm disc apart. Key words: Staphylococcus aureus, methicillin resistant S. aureus (MRSA), methicillin sensitive S. aureus (MSSA), inducible macrolide-lincosamide-streptogramin B phenotype (iMLSB), cMLSB, D-test.