Evaluation of the XpertMTB/RIF for the Diagnosis of Pulmonary Tuberculosis Among the Patients Attending DOTS Center Parsa District of Nepal

Tuberculosis diagnosis and monitoring rely in Sputum microscopy of National Tuberculosis Programme, Nepal because of its low cost and easier to perform. Direct sputum microscopy is popular worldwide. Currently, there are 533 microscopy centres catering for sputum microscopy examination throughout the country. Most of the microscopy centres are established within government jurisdiction and remaining are established as non-governmental organization as well as private sectors.A cross-sectional study was conducted from July 2013 to January 2015. A total of 2091 patients were enrolled in the study who were attending the DOTS Centre in Parsa District of Public Health Office, Nepal. Smears stained with ZN stain methods were examined microscopically followed by the GeneXpert MTB/RIF assay.Out of 2091 suspected pulmonary TB patients enrolled for sputum microscopy and GeneXpert MTB/RIF for the confirmation of TB, the 1301(62.21%) were male and 790 (37.78%) were female. The maximum TB cases were from Parsa district (555, 26.5%). The comparative study of different diagnostic tools reveals the sensitivity of MTB/RIF was 95.50% (91.87, 97.82) and significantly higher than smear microscopy performed on the same fluid, which had a sensitivity of 61.97% (55.41, 68.21). Five of 127 smear-negative cases had MTB/RIF-positive un-centrifuged sputum, resulting in a specificity of 81.23% (75.95, 85.78), which was similar to smear microscopy 98.29 % (97.34, 98.97; p=0.121). The positive predictive value (PPV) and negative predictive value (NPV) of MTB/RIF were 96.85% (93.61, 98.72) and 94.95 % (93.52, 96.14), respectively. HIV co-infection did not impact sensitivity, specificity or liquid culture time to positivity (TTP). When MTB/RIF accuracy was evaluated using composite reference standard culture positivity from sputum, the sensitivity and specificity were similar to those obtained in the primary analysis using either definite TB versus possible and non-TB combined; definite and possible TB combined versus non-TB

The Dengue virus in Nepal: gaps in diagnosis and surveillance.

BACKGROUND: The introduction of the dengue virus (DENV) in Nepal is recent, first reports date back to 2004 from a Japanese traveller and limited information is available about DENV infection in the Nepali population. Within a decade after the first DENV detection, it is now endemic in multiple districts of Nepal with approximately 11.2 million people residing in the Terai belt being at risk of DENV infection. Sporadic cases of DENV infection have been reported every year for the past decade during the monsoon season, mainly in the Terai region. METHODS: Medline/Embase/Cochrane databases were reviewed for reports on the burden of dengue infection, diagnostic methods, and national surveillance. RESULTS: Four outbreaks were reported since 2004 including the diagnosis of all serotypes in 2006 and predominance of a single serotype in 2010 (DENV-1), 2013 (DENV-2), and 2016 (DENV-1). The clinical diagnoses showed a predominance of dengue fever while 4/917 (0.4%), 8/642 (1.2%) and 8/1615 (0.4%) dengue haemorrhagic fever/dengue shock syndrome cases were identified during the outbreaks in 2010, 2013 and 2016, respectively. The number of cases reported in males was significantly higher (67.4%) than in females. Disease occurrence was primarily found in the Terai region until 2010 and was increasingly detected in the Hilly region in 2016. CONCLUSION: In Nepal currently weak diagnostic facilities, very limited research on mosquitoes vectors, and poor surveillance of dengue leading to inappropriate detection and control of DENV. We surmise that improved basic research and epidemiological training courses for local scientists and laboratory personal at national and international level will help better understand the evolution and distribution of DENV transmission and its eventual control

Effect of lyophilization on infectivity and viral load of Adenovirus

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: 15-2

Pattern of Cancer in Nepal from 2003 to 2011

Correction: On 15th January 2017, the authors Sunil Kumar Sah and Naval Kishor Yadav were added to the author list.Cancer is global burden of disease in developed and developing countries. It is one of the main causes of death. The environmental factor and life styles are major causes of cancer.This hospital based retrospective study was carried out using data retrieved from the register maintained at seven cancer centers. The most common basis of diagnosis were microscopic (histopathological and cytopathological examination). The diagnosis was also based on clinical examination, radiological examination, endoscopy, biochemical and immunological tests.Most of the cancer cases were diagnosed at BPKMCH (23908) followed by BPKIHS (9668) and BH (5959) and few cases from KCH (518) in 2003 to 2011. The total number of cancer cases were increasing from 2003 to 2011 and it become double in 2011. Out of 75 district of Nepal, more number of cancer cases was found in Kathmandu, Sunsari, Morang, Chitwan, Lalitpur, Jhapa, Kaski, Nawalparasi, Rupendehi and Kavrepalchowk in 2010. Similarly, in 2011 more number of cancer cases was found in Kathmandu, Morang, Jhapa, Sunsari, Chitwan, Lalitpur, Rupendehi, Kaski, Saptari, Bhaktapur. Lung cancer was the common cancer and similarly, other prevalent cancers were cervical, breast, stomach, ovarian and colo-rectum cancer in 2003 to 2011. The common cancers were lung, cervical, breast, stomach, ovarian and colo-rectum. The number of patients is increasing, which may be due to change in life style and lack of education

Prevalence and risk of hepatitis e virus infection in the HIV population of Nepal

Background: Infection with the hepatitis E virus (HEV) can cause acute hepatitis in endemic areas in immune-competent hosts, as well as chronic infection in immune-compromised subjects in non-endemic areas. Most studies assessing HEV infection in HIV-infected populations have been performed in developed countries that are usually affected by HEV genotype 3. The objective of this study is to measure the prevalence and risk of acquiring HEV among HIV-infected individuals in Nepal. Methods: We prospectively evaluated 459 Human Immunodeficiency Virus (HIV)-positive individuals from Nepal, an endemic country for HEV, for seroprevalence of HEV and assessed risk factors associated with HEV infection. All individuals were on antiretroviral therapy and healthy blood donors were used as controls. Results: We found a high prevalence of HEV IgG (39.4%) and HEV IgM (15.3%) in HIV-positive subjects when compared to healthy HIV-negative controls: 9.5% and 4.4%, respectively (OR: 6.17, 95% CI 4.42-8.61, p < 0.001 and OR: 3.7, 95% CI 2.35-5.92, p < 0.001, respectively). Individuals residing in the Kathmandu area showed a significantly higher HEV IgG seroprevalance compared to individuals residing outside of Kathmandu (76.8% vs 11.1%, OR: 30.33, 95% CI 18.02-51.04, p = 0.001). Mean CD4 counts, HIV viral load and presence of hepatitis B surface antigen correlated with higher HEV IgM rate, while presence of hepatitis C antibody correlated with higher rate of HEV IgG in serum. Overall, individuals with HEV IgM positivity had higher levels of alanine aminotransferase (ALT) than IgM negative subjects, suggesting active acute infection. However, no specific symptoms for hepatitis were identified. Conclusions: HIV-positive subjects living in Kathmandu are at higher risk of acquiring HEV infection as compared to the general population and to HIV-positive subjects living outside Kathmandu

Hepatitis viruses in Kathmandu, Nepal: hospital-based study

Abstract Objective The objective of this study was to see the aetiology and outcome of sporadic acute viral hepatitis (AVH) in Kathmandu, Nepal. Results Among 210 patients, 94 (45%) were male and 116 (55%) were female. Mean age was 30 years. 52 (24.7%) out of 210 were positive for either of the hepatitis virus infection. Major causative agent for AVH among hepatitis positive patients were hepatitis E virus (HEV) in 36 (69.2%), followed by hepatitis A virus (HAV) 8 (15.3%), hepatitis B virus (HBV) 7 (13.4%) and hepatitis C virus (HCV) 1 (1.9%). The 158 (75.3%) patient were negative for all hepatitis viral markers. Co-infections with more than one virus were found in 4 (7.6%) patients. All liver-specific enzymes including bilirubin increased in hepatitis-infected patients. We found large number circulation of HEV in Kathmandu, Nepal, indicating that this region is endemic for hepatitis virus infection

Estimation of Transmission of Porphyromonas Gingivalis from Mother to Child through Saliva

Introduction: Chronic periodontitis is an infectious disease. Porphyromonas gingivalis is the major pathogen associated with it and can be found in all ecosystems in the oral cavity. The presence of this organism is highly correlated with preterm and low birth weight babies. So, this study aimed to assess vertical transmission of P.gingivalis from pregnant women to their new born. Methods: Forty six pregnant women with chronic periodontitis were recruited for this cross-sectional study. Whole unstimulated saliva was collected from them before delivery and from their new-borns within forty eight hours of birth. Quantification of P.gingivalis in the saliva samples was carried out by quantitative real time polymerase chain reaction. The obtained data were analysed by SPSS 16 program. Results: The results showed a significant correlation (P=0.002) between the number of P.gingivalis present in the mother’s saliva with that of the new-borns’ saliva. DNA copies of more than 5000/μl of P.gingivalis was found in 20 (43.5%) maternal saliva and 21 (45.7%) in new-borns’ saliva. Both Plaque index and Extent and Severity index showed no correlation (P>0.05) with DNA copies of P.gingivalis in new-borns’ saliva. Conclusions: The DNA copies of P.gingivalis found in new-borns’ saliva are in par with mother saliva, as the saliva sample obtained from new-borns’ were within forty eight hours of birth, no other environmental factor can have a direct role in its transmission. Thus, it can be concluded that P.gingivalis is vertically transmitted from mother to child

Profile of the 2016 dengue outbreak in Nepal

Abstract Objective The objective of this study was to obtain clinical, virological and demographic data detailing the 2016 dengue outbreak in Nepal. Results Dengue disease was first reported in Nepal in 2004 and several major outbreaks have occurred since then, with a significant impact on public health. An outbreak of dengue fever occurred in Nepal during June to November 2016, with a peak number of cases reported in September. 1473 patients with laboratory confirmed DENV infections visited or were admitted to hospitals during this period. The most common clinical symptoms included fever, headache, joint pain and thrombocytopenia. Serotyping of 75 serum samples from patients having fever for less than 4 days was carried out with a dengue virus (DENV) serotype-specific RT-PCR strategy. Our results indicate that the dengue outbreak in Nepal during 2016 was caused predominantly, if not exclusively, by DENV-1, representing a shift in the prevailing serotype from DENV-2, the dominant serotype characterizing the 2013 dengue epidemic in Nepal. Hopefully, this report will assist Nepalese public health agencies in developing improved dengue-related programs including mosquito-vector control, DENV surveillance, and diagnosis and treatment of dengue fever patients, in order to reduce the impact of future dengue epidemics