Prevalence of drug resistant Enterobacteriaceae in a Nepalese tertiary care hospital.

Antimicrobial resistance in Enterobacteriaceae is an emerging global public health problem. Numerous studies have reported community-acquired AmpC beta-lactamase and extended spectrum beta-lactamase (ESBL) producing Enterobacteriaceae in Nepal. However, there are limited data on community-acquired Metallo-beta-lactamase (MBL) producing Enterobacteriaceae. A hospital-based descriptive cross-sectional study was conducted using 294 Enterobacteriaceae isolates from a total of 2,345 different clinical specimens collected from patients attending a tertiary care hospital in Nepal. Bacteria were isolated using standard microbiological growth media and identified using biochemical tests. For antimicrobial susceptibility testing, Kirby-Bauer disc diffusion technique was used. AmpC, ESBL, and MBL productions were detected by using combined disc method. AmpC, ESBL, and MBL productions were detected in 19.4%, 29.6%, and 8.5% of total Enterobacteriaceae isolates respectively. Higher rates of beta-lactamases production were seen among the isolates from in-patients in comparison with those from out-patients. However, 11.6%, 25%, and 3.7% of the total isolates from out-patients were AmpC, ESBL, and MBL producers respectively. The co-production of the beta-lactamases was also detected, with two Klebsiella pneumoniae isolates producing all three beta-lactamases. One MBL producing Proteus vulgaris isolate that was pan-resistant with no remaining treatment options was also isolated. Prevalence of drug resistant Enterobacteriaceae in our study was very high. Detection of AmpC, ESBL, and MBL positive isolates from out-patients, who did not have recent history of hospital visit, indicated the community dissemination of the drug resistant bacteria. This is a matter of great concern and an immediate attention to formulate strategies to prevent further development and spread of antibiotic resistance is required.

Prevalence and correlates of Helicobacter pylori infection among under-five children, adolescent and non-pregnant women in Nepal: Further analysis of Nepal national micronutrient status survey 2016.

Most of the Helicobacter pylori infections occur in developing countries. The risk factors for H. pylori infections are poverty, overcrowding, and unhygienic conditions, which are common problems in under-privileged countries such as Nepal. Despite having a high risk of H. pylori infections, no national level study has been conducted to assess prevalence and correlates of H. pylori infection in Nepal. Therefore, we hypothesized that micronutrients such as iron, vitamin B12 deficiency, socio-economic status, and nutritional status correlate with the prevalence of H. pylori infection in Nepal. We studied prevalence and correlates of H. pylori infection among under-five children, adolescents aged 10-19 years and married non-pregnant women aged 20-49 years using data from the Nepal National Micronutrient Status Survey 2016 (NNMSS-2016). H. pylori infection was examined in stool of 6-59 months old children and 20-49 years old non-pregnant women whereas the rapid diagnostic kit using blood sample was used among adolescent boys and girls. Prevalence of H. pylori infection was 18.2% among 6-59 months old children, 14% among adolescent boys and 16% among adolescent girls aged 10-19 years; and 40% among 20-49 years non-pregnant women. Poor socioeconomic status, crowding, and unhygienic condition were found to be positively associated with higher incidence of H. pylori infections. No significant correlation was observed between nutritional and micronutrients status (iron or risk of folate deficiency) with H. pylori infection. Findings from this study suggest that poverty-associated markers are primary contributors of H. pylori infections in Nepalese communities. To control acquisition and persistence of H. pylori infection in Nepal, we suggest improved management of safe drinking water and implementation of sanitation and hygiene programs, with a focus on those of lower socioeconomic status.

Extended spectrum ß-lactamase producing uropathogenic Escherichia coli and the correlation of biofilm with antibiotics resistance in Nepal.

BACKGROUND: Urinary tract infection (UTI) is one of the frequently diagnosed infectious diseases which is caused mainly by Escherichia coli. E. coli confers resistance against the two major classes of antibiotics due to the production of extended spectrum ß-lactamase enzymes (ESBL), biofilm, etc. Biofilm produced by uropathogenic E. coli (UPEC) protects from host immune system and prevent entry of antimicrobial compounds. The main objective of this cross-sectional study was to determine the correlation of biofilm production and antibiotic resistance as well as to characterize the pgaA and pgaC genes responsible for biofilm formation among uropathogenic ESBL producing E. coli. METHODS: A total of 1977 mid-stream urine samples were examined and cultured for bacterial strain identification. ESBL was detected by combined disc method following CLSI whereas biofilm formation was analyzed by semi-quantitative method. Furthermore, the pgaA and pgaC genes responsible for biofilm formation in UPEC were detected by multiplex PCR. All the statistical analyses were done via IBM SPSS Statistics 21 where Pearson's correlation test were used to determine correlation (-1 ≥ r ≤ 1). RESULTS: E. coli was the predominant causative agent, which accounted 159 (59.3%) of the Gram-negative bacteria, where 81 (50.9%) E. coli strains were found to be ESBL producers. In addition, 86 (54.1%) E. coli strains were found to be biofilm producers. Both the pgaA and pgaC genes were detected in 45 (93.7%) the UPEC isolates, which were both biofilm and ESBL producers. Moreover, there was a positive correlation between biofilm and ESBL production. CONCLUSION: The analyses presented weak positive correlation between biofilm and ESBL production in which biofilm producing UPEC harbors both pgaA and pgaC genes responsible for biofilm formation.

Biofilm and metallo beta-lactamase production among the strains of Pseudomonas aeruginosa and Acinetobacter spp. at a Tertiary Care Hospital in Kathmandu, Nepal.

INTRODUCTION: Pseudomonas aeruginosa and Acinetobacter spp. are found to be associated with biofilm and metallo-ß-lactamase production and are the common causes of serious infections mainly in hospitalized patients. So, the main aims of this study were to determine the rates of biofilm production and metallo beta-lactamase production (MBL) among the strains of Pseudomonas aeruginosa and Acinetobacter spp. isolated from hospitalized patients. METHODS: A total of 85 P. aeruginosa isolates and 50 Acinetobacter spp. isolates isolated from different clinical specimens from patients admitted to Shree Birendra Hospital, Kathmandu, Nepal from July 2013 to May 2014 were included in this study. The bacterial isolates were identified with the help of biochemical tests. Modified Kirby-Bauer disc diffusion technique was used for antimicrobial susceptibility testing. Combined disc diffusion technique was used for the detection of MBL production, while Congo red agar method and tube adherence method were used for detection of biofilm production. RESULTS: Around 16.4% of P. aeruginosa isolates and 22% of the strains of Acinetobacter spp. were metallo ß-lactamase producers. Out of 85 P. aeruginosa isolates, 23 (27.05%) were biofilm producers according to tube adherence test while, only 13 (15.29%) were biofilm producers as per Congo red agar method. Similarly, out of 50 Acinetobacter spp. 7 (14%) isolates were biofilm producers on the basis of tube adherence test, while only 5 (10%) were positive for biofilm production by Congo red agar method. Highest rates of susceptibility of P. aeruginosa as well as Acinetobacter spp. were seen toward colistin. CONCLUSION: In our study, biofilm production and metallo beta-lactamase production were observed among Pseudomonas aeruginosa and Acinetobacter spp. However, no statistically significant association could be established between biofilm production and metallo beta-lactamase production.

Isolation of Multidrug Resistant Bacteria from Patients Medical Charts.

BACKGROUND: Patient's medical charts in hospitals are potentially contaminated by pathogenic bacteria and might act as vehicles for transmission of bacterial infections.This study was aimed to determine the rate of contamination of medical charts by multidrug resistant bacteria. METHODS: Sampling of total 250 patient's medical charts from different wards was done with the help of cotton swabs soaked in sterile normal saline. The swabs thus collected were cultured using standard microbiological procedures.The colonies grown were then identified with the help of colony morphology, Gram's stain and biochemical tests. Antimicrobial susceptibility testing was performed by using Kirby-Bauer disc diffusion technique. RESULTS: Of the total 250 charts sampled, 98.8% grew bacteria; Bacillus spp. in 40.7%, followed by Staphylococcus aureus (17%), coagulase-negative Staphylococcus spp.(CoNS) (17%), Citrobacter freundii (9.6%) and Acinetobacter spp. (4.5%). Rate of multidrug resistance was highest in Acinetobacter spp. (50%). Among 83 isolates of S. aureus, methicillin resistance was found in 29 isolates. Similarly, two out of total 9 isolates of Enterococcus spp. were vancomycin resistant. CONCLUSIONS: This study showed that patient's medical charts were contaminated with multidrug resistant bacteria including methicillin resistant S. aureus and vancomycin resistant Enterococcus spp. Strict hand washing before and after handling medical charts is recommended.

Extended spectrum beta-lactamase and metallo beta-lactamase production among Escherichia coli and Klebsiella pneumoniae isolated from different clinical samples in a tertiary care hospital in Kathmandu, Nepal.

BACKGROUND: Extended spectrum beta-lactamase (ESBL) and metallo beta-lactamase (MBL) production in Klebsiella pneumoniae and Escherichia coli are the commonest modes of drug resistance among these commonly isolated bacteria from clinical specimens. So the main purpose of our study was to determine the burden of ESBL and MBL production in E. coli and K. pneumoniae isolated from clinical samples. Further, the antimicrobial susceptibility patterns of E. coli and K. pneumoniae were also determined. METHODS: A cross-sectional study was conducted at Om Hospital and Research Centre, Kathmandu, Nepal by using the E. coli and K. pneumoniae isolated from different clinical samples (urine, pus, body fluids, sputum, blood) from May 2015 to December 2015. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Extended spectrum beta-lactamase production was detected by combined disc method using ceftazidime and ceftazidime/clavulanic acid discs and cefotaxime and cefotaxime/clavulanic acid discs. Similarly, metallo beta-lactamase production was detected by combined disc assay using imipenem and imipenem/ethylenediaminetetracetate discs. Bacteria showing resistance to at least three different classes of antibiotics were considered multidrug resistant (MDR). RESULTS: Of total 1568 different clinical samples processed, 268 (17.1%) samples were culture positive. Among which, E. coli and K. pneumoniae were isolated from 138 (51.5%) and 39 (14.6%) samples respectively. Of the total isolates 61 (34.5%) were ESBL producers and 7 (4%) isolates were found to be MBL producers. High rates of ESBL production (35.9%) was noted among the clinical isolates from outpatients, however no MBL producing strains were isolated from outpatients. Among 138 E. coli and 39 K. pneumoniae, 73 (52.9%) E. coli and 23 (59%) K. pneumoniae were multidrug resistant. The lowest rates of resistance was seen toward imipenem followed by piperacillin/tazobactam, amikacin and cefoperazone/sulbactam. CONCLUSIONS: High rate of ESBL production was found in the E. coli and K. pneumoniae isolated from outpatients suggesting the dissemination of ESBL producing isolates in community. This is very serious issue and can't be neglected. Regular monitoring of rates of ESBL and MBL production along with multidrug resistance among clinical isolates is very necessary.

Use of Genotype MTBDRplus Assay for Diagnosis of Multidrug-Resistant Tuberculosis in Nepal.

The main aims of this study were to study the patterns of mutations in rpoB, katG, and inhA genes in Mycobacterium tuberculosis strains isolated from patients from Nepal and to evaluate the performance of genotype MTBDRplus assay, taking conventional drug susceptibility testing as gold standard for diagnosis of MDR-TB. A total of 69 Mycobacterium tuberculosis strains isolated from 73 smear positive sputum samples from patients suspected of suffering from multidrug-resistant tuberculosis were used in our study. The drug susceptibility pattern of Mycobacterium tuberculosis isolated from these sputum specimens was determined by using genotype MTBDRplus assay taking conventional drug susceptibility testing as reference. The sensitivity and specificity of the genotype MTBDRplus assay for the detection of MDR-TB were found to be 88.7% and 100%, respectively. 88.7% of the rifampicin resistant isolates had mutations in rpoB gene. Similarly, 79.7% and 9.4% of isoniazid resistant isolates had mutations in katG and inhA genes, respectively. Genotype MTBDRplus assay was found to be very rapid and highly sensitive and specific method for diagnosis of MDR-TB and will be very helpful for early diagnosis of MDR-TB in high tuberculosis burden countries.

AmpC and extended spectrum beta-lactamases production among urinary isolates from a tertiary care hospital in Lalitpur, Nepal.

BACKGROUND: Production of AmpC and extended spectrum beta-lactamases among urinary isolates has created a serious problem to the successful management of the urinary tract infection. The main purpose of this study was to determine the rates of the extended spectrum beta-lactamase (ESBL) production and AmpC beta-lactamase (ABL) production among urinary isolates. RESULTS: Among total 564 urinary isolates, 514 (91.1%) were gram negative bacilli and 50 (8.9%) were gram positive cocci. E. coli (76.1%) was the most common bacteria isolated. Staphylococcus aureus (6.7%) was the predominant gram positive bacteria isolated. 35 (6.8%) of the 514 gram negative bacilli were ESBL producers. Similarly, 14 (2.7%) of the gram negative bacilli were ABL producers. Only one isolate was ESBL and ABL co-producer. Highest rate of susceptibility of gram negative bacteria was seen toward amikacin (97.3%) followed by imipenem (94.4%). Similarly, highest rate of susceptibility among gram positive cocci was seen toward vancomycin (100%) followed by amikacin (93.5%). CONCLUSIONS: Low rates of AmpC and extended spectrum beta-lactamases production in comparison to other previous studies were reported. On the basis of the antimicrobial susceptibility patterns of the bacteria we reported in our study, amikacin, imipenem and nitrofurantoin can be used for the preliminary treatment of urinary tract infections caused by gram negative bacteria and vancomycin and amikacin for treatment of urinary tract infections caused by gram positive bacteria.

Phytochemical screening and study of antioxidant, antimicrobial, antidiabetic, anti-inflammatory and analgesic activities of extracts from stem wood of Pterocarpus marsupium Roxburgh.

AIMS: The main aims of the study were to evaluate the phytochemical constituents and to study the antioxidant, antimicrobial, antidiabetic, anti-inflammatory, and analgesic activities of extracts from stem wood of Pterocarpus marsupium. MATERIALS AND METHODS: Ethanol, acetone and isopropyl alcohol (IPA) (1:1) extracts of stem wood of P. marsupium were subjected to phytochemical screening and analysis of biological activities from August 2015 to January 2016. The antioxidant assay was carried out using 2, 2-diphenyl-1-picrylhydrazyl scavenging method, antimicrobial activity testing by cup diffusion method, antidiabetic test evaluation by oral glucose tolerance test in mice, anti-inflammatory effect was evaluated by hind paw edema method in mice and analgesic test evaluation by a chemical writhing method in mice. RESULTS: The results of the study revealed that P. marsupium is a source of various phytoconstituents such as alkaloids, glycosides, saponins, tannins, proteins, carbohydrates, cardiac glycosides, flavonoids, and terpenoids. Both the acetone and IPA extract as well as the ethanol extract of stem wood of P. marsupium exhibited a dose-dependent antioxidant activity. Acetone and IPA extract showed antibacterial activity against Gram-positive bacteria, while the ethanolic extract was found to possess antidiabetic activity. The antidiabetic activity of the extract was found to be time and dose-dependent. Similarly, the acetone and IPA extract was found to have anti-inflammatory activity, which was also time and dose-dependent. Furthermore, the ethanolic extract showed analgesic activity, which was dose-dependent. The ethanolic extract was found to be nontoxic. CONCLUSIONS: Thus, this study laid sufficient background for the further research on extracts from stem wood of P. marsupium for identification, subsequent purification and isolation of compounds having antibacterial, antidiabetic, anti-inflammatory, and analgesic activities.

Bacteremia and Urinary Tract Infection Caused by Chromobacterium violaceum: Case Reports from a Tertiary Care Hospital in Kathmandu, Nepal.

Chromobacterium violaceum is ubiquitous in the environment of tropical and subtropical regions but the infections caused by this organism are rare and the urinary tract infections caused by it are even rarer. Due to the propensity for hematogenous spread leading to fatal sepsis, the infections caused by Chromobacterium violaceum have high mortality rate (65-80%) with death occurring in as less as one week of acquiring infection. So, prompt proper treatment is necessary for successful treatment of the infections but, due to the rarity of the infections caused by the organism, there is limited awareness among the clinicians regarding the infections caused by this organism. Here, we reported a case of urinary tract infection caused by Chromobacterium violaceum in a 84-year-old male, who was a kidney patient, and another case of bacteremia caused by the same bacterium in a road traffic accident patient (22-year-old male), both of which were managed with the timely suitable treatment.

Bacteriological Profile and Antimicrobial Susceptibility Patterns of Bacteria Isolated from Pus/Wound Swab Samples from Children Attending a Tertiary Care Hospital in Kathmandu, Nepal.

In Nepal, little is known about the microbiological profile of wound infections in children and their antimicrobial susceptibility patterns. Total of 450 pus/wound swab samples collected were cultured using standard microbiological techniques and the colonies grown were identified with the help of biochemical tests. The antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Methicillin-resistant Staphylococcus aureus isolates were detected by using cefoxitin disc and confirmed by determining minimum inhibitory concentrations (MIC) of oxacillin. 264 (59%) samples were culture positive. The highest incidence of bacterial infections was noted in the age group of less than 1 year (76%). Out of 264 growth positive samples, Gram-positive bacteria were isolated from 162 (61%) samples and Gram-negative bacteria were found in 102 (39%) samples. Staphylococcus aureus (99%) was the predominant Gram-positive bacteria isolated and Pseudomonas aeruginosa (44%) was predominant Gram-negative bacteria. About 19% of S. aureus isolates were found to be methicillin-resistant MIC of oxacillin ranging from 4 µg/mL to 128 µg/mL. Among the children of Nepal, those of age less than 1 year were at higher risk of wound infections by bacteria. Staphylococcus aureus followed by Pseudomonas aeruginosa were the most common bacteria causing wound infections in children.

Anti Japanese encephalitis virus IgM positivity among patients with acute encephalitic syndrome admitted to different hospitals from all over Nepal.

The Japanese encephalitis virus (JEV) infection is one of the major public health problems in Nepal because of its increasing disease morbidity and mortality. The main purpose of this study was to determine the anti-JEV IgM positivity among acute encephalitis syndromic cases from all over Nepal. The present study was conducted at National Public Health Laboratory, Kathmandu, Nepal from April 2015 to October 2015. A total of 671 (418 CSF and 253 serum) samples were collected from 625 patients with acute encephalitic syndrome, admitted to different hospitals from all over Nepal. IgM antibody capture enzyme linked immunosorbent assay (ELISA) was used for the detection of anti-JEV IgM positive cases. The rate of anti-JEV IgM positivity was found to be 21.12%. The majority of positive cases (50%) were from the age group below 15 years, with the highest numbers of cases occurring in September (55.30%). Among all the anti-JEV IgM positive cases, higher numbers of cases were males. Geographically, the highest numbers of anti-JEV IgM positive cases were recorded from Terai region. Similarly, largest numbers of anti-JEV IgM positive cases were reported from Kailai district followed by those from Kanchanpur. However, anti-JEV IgM positive cases were also reported from hill districts. Continuation of active surveillance and vector control measures, proper management of diagnostic facilities and expanded program of immunization in JE endemic areas should be strongly emphasized to reduce the endemicity of the disease.

Antibiotic resistance and biofilm production among the strains of Staphylococcus aureus isolated from pus/wound swab samples in a tertiary care hospital in Nepal.

BACKGROUND: The increasing drug resistance along with inducible clindamycin resistance, methicillin resistance and biofilm production among the strains of Staphylococcus aureus are present as the serious problems to the successful treatment of the infections caused by S. aureus. So, the main objectives of this study were to determine the antimicrobial susceptibility patterns along with the rates of inducible clindamycin resistance, methicillin resistance and biofilm production among the strains of S. aureus isolated from pus/wound swab samples. METHODS: A total of 830 non-repeated pus/wound swab samples were processed using standard microbiological techniques. The colonies grown were identified on the basis of colony morphology, Gram's stain and biochemical tests. Antimicrobial susceptibility testing was performed by Kirby-Bauer disc diffusion technique. Detection of inducible clindamycin resistance was performed by D test, while detection of methicillin resistant S. aureus (MRSA) was performed by determination of minimum inhibitory concentration of oxacillin by agar dilution method. Similarly, detection of biofilm formation was performed by microtiter plate method. Strains showing resistance to three or more than three different classes of antibiotics were considered multidrug resistant. RESULTS: Total 76 samples showed the growth of S. aureus, among which 36 (47.4%) contained MRSA and 17 (22.4%) samples were found to have S. aureus showing inducible clindamycin resistance. Among the S. aureus isolated from outpatients, 41.9% were MRSA. Highest rates of susceptibility of S. aureus were seen toward linezolid (100%) and vancomycin (100%). Similarly, S. aureus isolated from 35 (46.1%) samples were found to be biofilm producers. Higher rate of inducible clindamycin resistance was seen among MRSA in comparison to methicillin susceptible S. aureus (MSSA). Similarly, higher rates of multidrug resistance and methicillin resistance were found among biofilm producing strains in comparison to biofilm non producing strains. CONCLUSIONS: The rate of isolation of MRSA from community acquired infections was found to be high in Nepal. Increased rate of inducible clindamycin resistance as compared to previous studies in Nepal was noted. So for the proper management of the infections caused by S. aureus, D test for the detection of inducible clindamycin resistance should be included in the routine laboratory diagnosis. Further, detection of biofilm production should also be included in the routine tests. Linezolid and vancomycin can be used for the preliminary treatment of the serious infections caused by S. aureus.

Detection of Methicillin Resistant Staphylococcus aureus and Determination of Minimum Inhibitory Concentration of Vancomycin for Staphylococcus aureus Isolated from Pus/Wound Swab Samples of the Patients Attending a Tertiary Care Hospital in Kathmandu, Nepal.

The present study was conducted to evaluate the performance of cefoxitin disc diffusion method and oxacillin broth microdilution method for detection of methicillin resistant S. aureus (MRSA), taking presence of mecA gene as reference. In addition, inducible clindamycin resistance and beta-lactamase production were studied and minimum inhibitory concentration (MIC) of vancomycin for S. aureus isolates was determined. A total of 711 nonrepeated pus/wound swab samples from different anatomic locations were included in the study. The Staphylococcus aureus was identified on the basis of colony morphology, Gram's stain, and biochemical tests. A total of 110 (15.47%) S. aureus isolates were recovered, of which 39 (35.50%) isolates were identified as MRSA by cefoxitin disc diffusion method. By oxacillin broth microdilution method, 31.82% of the Staphylococcus aureus isolates were found to be MRSA. However, mecA gene was present in only 29.1% of the isolates. Further, beta-lactamase production was observed in 71.82% of the isolates, while inducible clindamycin resistance was found in 10% of S. aureus isolates. The MIC value of vancomycin for S. aureus ranged from 0.016 µg/mL to 1 µg/mL. On the basis of the absolute sensitivity (100%), both phenotypic methods could be employed for routine diagnosis of MRSA in clinical microbiology laboratory; however cefoxitin disc diffusion could be preferred over MIC method considering time and labour factor.

Prevalence of dengue and diversity of cultivable bacteria in vector Aedes aegypti (L.) from two dengue endemic districts, Kanchanpur and Parsa of Nepal.

BACKGROUND: Dengue fever, an endemic arboviral disease, represents one of the major public health concerns in Nepal. It is transmitted by bites of infected Aedes aegypti and Aedes albopictus, the former being primary vector. The bacterial community plays a significant role in biology of mosquitoes; however, the bacterial communities of primary vector A. aegypti remain unstudied in Nepal. The study was designed to determine the rate of dengue seropositivity and to explore the bacterial diversity of A. aegypti from dengue endemic districts, Kanchanpur and Parsa of Nepal. METHODS: A cross-sectional study was conducted between June 2013 and November 2013 at two hospitals of Kanchanpur and Parsa. A total of 221 serum samples were collected from patients (inpatients and outpatients) suspected of suffering from dengue fever and attending Mahakali Zonal Hospital, Mahendranagar, Kanchanpur, and Narayani Zonal Hospital, Birgunj, Parsa. Detection of anti-dengue IgM was performed by using human dengue IgM capture ELISA. The larvae and pupae of mosquitoes (A. aegypti) were collected, reared, and emerged. Then, the bacteria were isolated and identified from the gut of identified mosquitoes by using standard methods. RESULTS: Out of total 221 serum samples collected from patients suspected of suffering from dengue fever, 34 (15.38%) were positive for anti-dengue IgM. Gram-negative bacteria were isolated in largest proportion (63%) followed by gram-positive cocci (23.27%) and gram-positive rods (13.73%). The most common cultivable bacteria isolated were Staphylococcus spp., Pseudomonas spp., and Acinetobacter spp. The average bacterial load in the vectors was 3.91 × 104 cfu/ml. CONCLUSIONS: High rate of anti-dengue IgM seropositivity was reported in our study. The environmental bacteria were predominantly isolated and identified in A. aegypti. The paratransgenic approach to control vector might be possible by spreading the genetically modified bacteria in larval habitat or shelter of adult mosquitoes.

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.

Prevalence of enteropathogens in children under 15 years of age with special reference to parasites in Kathmandu, Nepal; a cross sectional study.

In developing countries like Nepal, gastrointestinal infections due to various parasites are common causes of morbidity and mortality in children. Present study was carried out from June 2013 to December 2013, among the children (<15 years of age) of Kathmandu Valley. Stool samples were collected from total 600 children (350 from four public schools and slum areas of Kathmandu valley and 250 from pediatric department of Tribhuvan University Teaching Hospital). The main objectives of this study were to investigate the intestinal parasitic infections in children below 15 years of age and their risk factors. However, some bacterial pathogens were also investigated. The overall prevalence of parasitic infections was 29.5 %. The rate of parasitic infections in children from community (39.43 %) was higher than that from hospital (15.6 %; p < 0.05). Giardia lamblia was the most common protozoan found to be causing infections in children and among helminths Ascaris lumbricoides was the most common worm isolated. Higher rates of parasitic infections were found in children of illiterate parents (38.17 %), children using untreated drinking water (49.77 %) and children having habit of consuming raw vegetables (31.50 %; p < 0.05). The present study indicated that the rate of infections due to enteropathogenic parasites was high among children of Nepal.

Drug susceptibility patterns of the Mycobacterium tuberculosis isolated from previously treated and new cases of pulmonary tuberculosis at German-Nepal tuberculosis project laboratory, Kathmandu, Nepal.

BACKGROUND: Multidrug resistant tuberculosis (MDR-TB) is a serious public health problem in Nepal. It is a major obstacle for the control of the tuberculosis. The main objectives of this study were to determine the prevalence of the multidrug resistant pulmonary tuberculosis and to evaluate the drug susceptibility patterns of Mycobacterium tuberculosis isolated from previously treated and newly diagnosed cases of pulmonary tuberculosis. METHODS: A cross-sectional study was conducted from March 2013 to August 2013 at German-Nepal tuberculosis project (GENETUP) laboratory, Kathmandu, Nepal. For this the sputum samples from total of 153 (49 new and 104 previously treated) suspected pulmonary tuberculosis patients were used. The diagnosis of the tuberculosis was performed by using fluorescent microscopy and culture, while the drug susceptibility testing of Mycobacterium tuberculosis was performed by proportion method. Lowenstein-Jensen (L-J) medium was used for the culture of Mycobacterium tuberculosis and the colonies grown were identified on the basis of the colony morphology, pigment production and biochemical characteristics. RESULTS: The prevalence of MDR-TB among all the cases of culture positive pulmonary tuberculosis was 15.6 %. The rate of MDR-TB among previously treated culture positive tuberculosis patients was 19.4 % and that among newly diagnosed culture positive pulmonary tuberculosis cases was 7.1 %. The highest rate of resistance of Mycobacterium tuberculosis, was toward streptomycin (24.4 %) followed by isoniazid (23 %), rifampicin (17.8 %) and ethambutol (15.6 %). Among the total of MDR-TB cases among previously treated patients, highest percentage of the cases were relapse (61.1 %) followed by chronic (16.7 %). CONCLUSIONS: The high prevalence of DR/MDR-TB in our study reflects poor implementation of tuberculosis control program. On the basis of the drug susceptibility patterns of M. tuberculosis we found in our study, we recommend to include ethambutol instead of streptomycin in the multidrug therapy for the treatment of tuberculosis patients in Nepal. Further, due to high rate of MDR-TB among previously treated patients, we do not recommend to use first line drugs for the treatment of pulmonary tuberculosis among previously treated patients.