The Double Burden of COVID-19 and Dengue in Nepal: The challenges ahead.

Coronavirus disease 2019 (COVID-19) pandemic has caused significant impact on the health care system. As a consequence, diagnosis and treatment of vector borne diseases including dengue has been equally affected. Nepal is no exception to this, where COVID-19 cases is exponentially increased and all resources are concentrated on its prevention, control and management. Dengue, one of the major vector-borne diseases in Nepal, is apparently overlooked despite approaching the peak season of the disease. The aim of this paper is to describe the double burden of COVID-19 and dengue in Nepal, particularly highlighting the co-circulation and possible coinfections. This has posed higher risk of increased severity, more severe cases and deaths in Nepal. Moreover, potential misdiagnosis of these viral diseases may lead to delayed or, inappropriate treatment and poor allocation of resources.

Impact of intensified case-finding strategies on childhood TB case registration in Nepal.

SETTING: Seven intervention districts with intensified childhood tuberculosis (TB) case-finding strategies implemented by a non-governmental organisation and seven control districts under the National Tuberculosis Programme, Nepal. OBJECTIVES: To assess the differences in childhood TB case registrations and case registration rates per 100 000 population between two time periods (Year 1 = March 2012-March 2013 and Year 2 = March 2013-March 2014) in intervention and control districts. DESIGN: Retrospective record review using routinely collected data. RESULTS: Childhood TB cases increased from 271 to 360 between Years 1 and 2 in the intervention districts (case registration rate from 18.2 to 24.2/100 000) and from 97 to 113 in the control districts (13.4 to 15.6/100 000): the increases were significantly higher in the intervention districts compared with the control districts. The increases were also significantly higher in children aged 0-4 years and in those with smear-negative pulmonary TB and extra-pulmonary TB. Of the various case-finding strategies, household contact screening, private-public mix services and mobile health chest camps produced the highest yield of TB. CONCLUSION: A package of intensified case-finding strategies in children was associated with an increase in childhood TB case registrations in Nepal. Additional diagnostic approaches to increase case registrations also need to be considered.

Contexte : Sept districts d'intervention avec des stratégies intensifiées de recherche active des cas de tuberculose (TB) mis en œuvre par une organisation non gouvernementale et sept districts témoins gérés par le Programme National Tuberculose au Népal.Objectifs : Evaluer les différences en termes d'enregistrement des cas de TB de l'enfant et de taux d'enregistrement pour 100 000 population entre deux périodes (année 1 = mars 2012 à mars 2013 et année 2 = mars 2013 à mars 2014) dans les districts d'intervention et les districts témoins.Schéma : Revue de dossiers rétrospective grâce aux données recueillies en routine.Résultats : Les cas de TB de l'enfant ont augmenté de 271 à 360 entre l'année 1 et l'année 2 dans les districts d'intervention (le taux d'enregistrement est passé de 18,2 à 24,2/100 000) et de 97 à 113 dans les districts témoins (13,4 à 15,6/100 000) : les augmentations ont été significativement plus importantes dans les districts d'intervention par rapport aux districts témoins. Les augmentations ont également été plus importantes chez les enfants de 0 à 4 ans et chez ceux qui ont eu une TB pulmonaire à frottis négatif et extra-pulmonaire. Parmi diverses stratégies de recherche des cas, le dépistage des contacts familiaux, les services conjoints privés-publics et les camps de santé mobiles pour la TB ont été les plus performants.Conclusion : Un paquet de stratégies intensifiées de recherche des cas a été associé à une augmentation des enregistrements de cas de TB de l'enfant au Népal. Il faut également envisager des approches diagnostiques supplémentaires pour augmenter encore l'enregistrement des cas.

Marco de referencia: Siete distritos de intervención en los cuales una organización no gubernamental aplica estrategias de búsqueda intensificada de casos de tuberculosis (TB) en los niños y siete distritos testigos del Programa Nacional contra la Tuberculosis de Nepal.Objetivos: Evaluar las diferencias en el registro de los casos de TB en la niñez y la tasa de registro de TB por 100 000 habitantes, en dos períodos (el primer año, de marzo del 2012 a marzo del 2013 y el segundo, de marzo del 2013 a marzo del 2014) en los distritos de intervención y los distritos testigos.Método: Fue este un estudio retrospectivo en el cual se analizaron los datos recogidos de manera sistemática.Resultados: Los casos de TB en la niñez aumentaron de 271 a 360 durante los períodos del estudio en los distritos de intervención (tasa de registro de casos de 18,2 a 24,2/100 000) y de 97 a 113 en los distritos testigos (de 13,4 a 15,6/100 000) y el incremento de los casos fue significativamente mayor en los distritos de intervención. Se observó un aumento con significación estadística en los niños del grupo de 0 a 4 años de edad, en los niños con TB pulmonar y baciloscopia negativa y con TB extrapulmonar. De las diferentes estrategias de búsqueda de casos, la detección sistemática de los contactos domiciliarios, los servicios mixtos público y privado y los puestos móviles de campaña de salud respiratoria alcanzaron el mayor rendimiento diagnóstico de TB.Conclusión: La introducción de un conjunto de estrategias de búsqueda intensificada de casos en los niños se asoció con un aumento en el registro de los casos de TB en Nepal. Se precisa también examinar la posibilidad de aplicar nuevos enfoques diagnósticos con el propósito de mejorar el registro de casos.

In vitro Temocillin efficacy against extended spectrum ß-lactamase producing multidrug resistant gram negative bacterial isolates from Nepal.

Temocillin is relatively more stable against most ß-lactamases and requires re-evaluation to include it in common clinical practice as a therapeutic alternative. At the National Reference Laboratory of Nepal, we evaluated multidrug resistance (MDR) and extended spectrum ß-lactamase (ESBL) phenotypes among 292 gram-negative clinical bacterial isolates of 18 different genera during 2009/2010 by Kirby-Bauer disc diffusion method following CLSI guidelines. ESBL screen positive isolates were tested for Temocillin efficacy by disc diffusion method following British Society of Antimicrobial Chemotherapy (BSAC) guidelines and other antibiotics following Clinical and Laboratory Standards Institute (CLSI) guidelines. Of the 292 isolates, 75.0% isolates were MDR, among which 61.6% were primarily screened positive for ESBL production but only 38.8% were confirmed as ESBL producers. We report relatively lower Temocillin resistance of 28.9% and 15.6% among MDR and ESBL positive populations, respectively. Among ESBL positive isolates, no Proteus mirabilis, 19.7% Escherichia coli and 33.3% Klebsiella oxytoca showed resistance to Temocillin, although such resistance was higher among Acinetobacter spp. (66.7%) and K. pneumoniae 50.0%. Among ESBL negative isolates, none of the K. oxytoca and few (13.3%) Acinetobacter spp. were resistant to Temocillin, while all Citrobacter freundii, Pseudomonas aeruginosa (85.7%) and K. pneumoniae (66.7%) showed Temocillin resistance. Only 14.8% and 3.0% of total MDR isolates were resistant to Imipenem and Meropenem, respectively. However, Imipenem resistance was remarkably high (86.7%) among ESBL negative Acinetobacter spp. than Meropenem (13.3%). Temocillin showed comparable efficacy against MDR and ESBL producing bacterial isolates and could be a next therapeutic option.

Current fluoroquinolone susceptibility criteria for Salmonella needs re-evaluation.

BACKGROUND: Disc diffusion technique is the routine susceptibility testing procedure for isolates of enteric fever, the most common clinical diagnosis among febrile patients in Nepal. OBJECTIVE: To evaluated the current fluoroquinolones (FQs) susceptibility criteria and nalidixic acid screening test in Salmonella enterica serovar Typhi and Paratyphi A. METHODS: S. Typhi and Paratyphi A strains isolated from 443 suspected enteric fever patients visiting National Public Health Laboratory (NPHL) during April through October 2008 were analyzed. All isolates were confirmed by standard microbiological procedures including serotyping. Antibiotic susceptibility testing was performed by using Kirby Bauer disc diffusion method and Clinical and Laboratory Standards Institute (CLSI) approved interpretive criteria. Agar dilution method was used to determine Minimum Inhibitory Concentration (MIC) of ciprofloxacin, ofloxacin and nalidixic acid. RESULT: Out of 41 Salmonella isolates, 80.49% were nalidixic acid resistant, with S. Paratyphi A showing higher resistance rate (88.23%) compared to S. Typhi (75%). The difference in both MIC and zone diameter in nalidixic acid susceptible and nalidixic acid resistant isolates was found to be significant (P < 0.001) and decreased susceptibility to FQs was strongly correlated (sensitivity and specificity of 100%) with resistance to nalidixic acid. Regression analysis of MIC against zone diameter based on the current CLSI recommended guidelines suggests that accommodation of current susceptible and resistant MIC requires increase in the zone diameter of ciprofloxacin and ofloxacin. CONCLUSION: Before using these drugs for management of enteric fever, appropriate identification of Salmonella isolates with reduced susceptibility to FQs is essential to limit the possible treatment failure and development of highly resistant strains. The current FQs susceptibility break point criteria for Salmonella need re-evaluation.

Values of lymphocyte subsets in Nepalese healthy adult population.

INTRODUCTION: CD4 T lymphocyte is the most commonly used cellular marker in tracking Human Immunodeficiency Virus (HIV) infection progression and monitoring effect of antiretroviral therapy (ART). Due to lack of local reference values of CD4 and other T lymphocytes in Nepal, most clinical decisions are based on the reference ranges of western countries. METHODS: This study was conducted at three major hospitals/laboratory of central, eastern and western Nepal during November 2008 through July 2009. Using the predefined criteria, 602 (200, 202 and 200 from central, eastern and western regions respectively) healthy adult volunteers of age range 18-60 years were recruited with equal representation from each age group and sex. Blood specimens were screened for HIV following standard algorithm using ELISA and two rapid test kits based on different principles. Samples with discordant test-results were excluded. HIV sero-negative specimens were further analyzed for CD4, CD8, and CD4:CD8 ratio, and absolute lymphocyte count (ALC) by FACS count (Becton Dickinson, USA) and automated cell counter (BC-3000 Plus, Shenzhen Mindray Biomedical Electronics, Germany) respectively. RESULTS: The average value (mean± standard deviation) of CD4, CD8, CD4/CD8 ratio and ALC of Nepalese adult population were found to be 786 ± 248, 567±230, 1.52 ± 0.59 and 2712 ± 836 respectively. All four parameters but CD8 were significantly different with sex and females had relatively higher values. However, none of these parameters reported significant difference with age except the ALC. CONCLUSIONS: Nepalese healthy adult populations have significantly different T lymphocyte subsets compared to other countries. The present reference ranges of CD4 and other T lymphocytes may be used for any clinical purposes including classifying and monitoring disease status in HIV infected individuals, immune status evaluation, monitoring ART and accordingly making amendment in national HIV treatment guidelines in Nepal.

Extended spectrum â-lactamase producing multidrug resistant clinical bacterial isolates at National Public Health Laboratory, Nepal.

Extended Spectrum â-lactamase (ESBL) producing multidrug resistant bacteria complicate therapeutic management and limit treatment options. Therefore, detection of ESBL-producing multidrug resistant (MDR) pathogens has a paramount importance. Between April 2009 and January 2010, a prospective study was carried out in National Public Health Laboratory with an objective to determine the status of ESBL producing MDR bacterial isolates from different clinical samples. Identification of the isolates was done by standard microbiological techniques and antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method following Clinical and Laboratory Standard Institute (CLSI) guidelines. ESBL screening among MDR isolates was done using Ceftriaxone, Aztreonam, Cefotaxime, Ceftazidime and Cefpodoxime followed by confirmation using MASTDISCS ID ES2L Detection Discs (CPD10). Data analysis was done by SPSS 16 software. Of the 314 bacterial isolates from 1601 different clinical specimens, 199 (63.4%) were MDR. Cefotaxime was found the reliable screening agent for ESBL detection with sensitivity and positive predictive value of 98.6% and 76.4% respectively. Sixtey nine (62.7%) isolates of the 110 tested MDR isolates were ESBL positive with at least one of the Combined Disk (CD) Assays. Escherichia coli (80%) was the major ESBL producer followed by Klebsiella pneumoniae (5.8%). A statistically significant relationship was found between increasing spectrum of drug resistance and ESBL production (p<0.05). Thus it is concluded that a higher rate of ESBL production prevail among MDR clinical bacterial isolates underscoring the need for routine ESBL detection in clinical laboratories.

Diagnostic dilemma of the single screening test used in the diagnosis of syphilis in Nepal.

Syphilis screening by the nontreponemal rapid plasma reagin (RPR) test is not usually followed up by specific treponemal tests in most of the resource poor healthcare settings of Nepal. We analyzed serum specimens of 504 suspected syphilis cases at the immunology department of the national reference laboratory in Nepal during 2007-2009 using RPR test and Treponema pallidum hemagglutination assay (TPHA). In overall, 35.7% were positive by both methods (combination) while 13.1% were RPR positive and TPHA negative, 8.7% were positive by TPHA only and 42.5% were negative by both methods. Among the RPR reactive (n = 246), 73.2% were positive by TPHA. Non-specific agglutination in RPR testing was relatively higher (26.8%) compared to TPHA (19.6%). Although TPHA was found more specific than RPR test, either of the single tests produced inaccurate diagnosis. Since the single RPR testing for syphilis may yield false positive results, specific treponemal test should be routinely used as confirmatory test to rule out false RPR positive cases. More attention needs to be paid on formulation of strict policy on the implementation of the existing guidelines throughout the country to prevent misdiagnosis in syphilis with the use of single RPR test.

Salmonella enterica serovar Paratyphi A: an emerging cause of febrile illness in Nepal.

With an aim to evaluate the isolation rate and antibiotic susceptibility pattern in Salmonella enterica serovar Typhi and S. Paratyphi A, 656 blood samples collected from clinically diagnosed enteric fever patients at National Public Health Laboratory, Nepal during January through December 2008 were processed. Isolates were identified by standard microbiological procedures including serotyping. Antibiotic susceptibility testing was performed by disc diffusion method and minimum inhibitory concentration (MIC) to ciprofloxacin, ofloxacin and nalidixic acid was determined by agar dilution method following CLSI guidelines. Altogether 59 isolates of S. Typhi (49.15%) and S. Paratyphi A (50.85%) were recovered. A total of 80% isolates were resistant to nalidixic acid with S. Paratyphi A (93%) showing significantly higher resistance (P < 0.05) compared to S. Typhi (66%). The nalidixic acid resistant S. Paratyphi A strains required significantly higher MICs (P < 0.001) to quinolone (MIC expressed as mean +/- SD for nalidixic acid 477.87 +/- 87.02 microg/mL, ofloxacin 1.8 +/- 0.63 microg/mL, ciprofloxacin 0.62 +/- 0.3 microg/mL) compared with that of S. Typhi (nalidixic acid 173.18 +/- 72.03 microg/mL, ofloxacin 0.43 +/- 0.11 microg/mL, ciprofloxacin 0.25 microg/mL). Increased MIC of fluoroquinolone (FQ) is of particular concern in emerging strains of S. Paratyphi A as exposure to these drugs fuels up further development of full FQ resistant populations. Use of FQs as the first-line drugs for empirical therapy and management of enteric fever in areas where these strains are prevalent is questionable and requires an urgent review.

Resistotypes of Vibrio cholerae 01 Ogawa Biotype El Tor in Kathmandu, Nepal.

Cholera continued to be a major diarrheal illness in Nepal and antibiotic resistance has appeared as a serious problem in cholera management. The study aimed at analyzing the distribution pattern of the resistotypes (R-types) of Vibrio cholerae in the Kathmandu valley, Nepal. During June 2008 to January 2009, 210 diarrheal specimens received at National Public Health Laboratory from suspected cholera patients were subjected to standard bacteriological investigation including biotyping and serotyping. Antimicrobial susceptibility pattern of V. cholerae isolates was determined by Kirby Bauer disc diffusion method following CLSI guidelines. A total of 57 (27%) V. cholerae isolated were recovered, all of which belonged to 01 Ogawa Biotype EL Tor. Based on antibiogram, V. cholerae isolates in our study revealed three distinct R-types: R-type I, R-type II and R-type III. All three R types showed resistance to furazolidone, nalidixic acid and cotrimoxazole while sensitive to ciprofloxacin and tetracycline. Additional resistance to ampicillin and erythromycin was observed respectively in R-type II and III. Different R-types showed unique month wise variations (P < 0.05). Differentiation of V. cholerae strains into R-types is an important tool. In addition to direct patient management, it may have implication in identifying the source and spread of infection, and understanding the distribution pattern in a particular geographical region.

Asymptomatic throat carriage rate and antimicrobial resistance pattern of Streptococcus pyogenes in Nepalese school children.

BACKGROUND: Streptococcus pyogenes or Group A streptococcus (GAS) causes several suppurative and non suppurative infections. In addition to pharyngitis and skin infections, GAS are also the causative agent of post-streptococcal infection syndromes such as acute rheumatic fever (ARF) and post-streptococcal glumerulonephritis (PSG). GAS frequently colonises in the throat of an asymptomatic person. Pharyngeal carriage rates of GAS among healthy school children vary with geographical location and seasons. OBJECTIVES: We carried out this preliminary study to determine the throat carriage rate and antimicrobial resistance trend of Streptococcus pyogenes or Group A streptococcus (GAS) among the Nepalese school children. MATERIALS AND METHODS: Four schools situated at different locations of Kathmandu valley were included in the study. Throat swabs from 350 students of age group 5-15 years were collected, immediately transported to the laboratory and were processed for S. pyogenes following standard microbiological procedures. Antimicrobial susceptibility testing of the isolates was performed by Kirby Bauer disc diffusion method following CLSI guidelines. RESULTS: S. pyogenes was isolated from 10.9% (38/350) of the screened children. The GAS colonisation rate was statistically insignificant (P>0.05) with sex and age sub-groups, although the rate was slightly higher among girls and age sub-group 9-12 years. No significant difference in carrier rate was observed among different schools (P>0.05). All isolates were susceptible to azithromycin. No resistance was detected for penicillin and its derivative antibiotic ampicillin. Highest resistance rate was observed for cotrimoxazole (71.0%) followed by chloramphenicol (7.8%), ciprofloxacin (5.2%) and erythromycin (5.2%). CONCLUSION: Antibiotic resistant GAS isolated from asymptomatic Nepalese school children is a public health concern. When screened and appropriately treated with antibiotics, carriers can be prevented from spreading of streptococcal infections in the school environment and the community. Preventing cross infections would ultimately reduce the incidence of life-threatening sequelae which are debilitating and difficult to treat. It is recommended to conduct regular screening and GAS surveillance in schools, and maintain rational use of antibiotics to minimise GAS carriage/infections and resistance.