Search

We report a novel piece-wise isothermal nucleic acid test (PINAT) for diagnosing pathogen-associated RNA that embeds an exclusive DNA-mediated specific probing reaction with the backbone of an isothermal reverse-transcription cum amplification protocol as a unified single-step procedure. This single step sample-to-result test method has been seamlessly integrated in an inexpensive, scalable, pre-programmable and portable instrument, resulting in a generic platform technology for detecting nucleic acid from a wide variety of pathogens. The test exhibited high sensitivity and specificity of detection when assessed using 200 double-blind patient samples for detecting SARS-CoV-2 infection conducted by the Indian Council of Medical Research (ICMR), reporting a positive and negative percent agreement of 94.6% and 98% respectively. We also established its efficacy in detecting influenza-A infection, performing the diagnosis at the point of collection with uncompromised detection rigor. The envisaged trade-off between advanced laboratory-based procedures with the elegance of common rapid tests renders the innovation to be ideal for deployment in resource-limited settings towards catering the needs of the underserved.

BackgroundIndia has made major progress in improving control of visceral leishmaniasis (VL) in recent years, in part through shortening the time infectious patients remain untreated. Active case detection decreases the time from VL onset to diagnosis and treatment, but requires substantial human resources. Targeting approaches are therefore essential to feasibility.MethodsWe analyzed data from the Kala-azar Management Information System (KAMIS), using village-level VL cases over specific time intervals to predict risk in subsequent years. We also graphed the time between cases in villages and examined how these patterns track with village-level risk of additional cases across the range of cumulative village case-loads. Finally, we assessed the trade-off between ACD effort and yield.ResultsIn 2013, only 9.3% of all villages reported VL cases; this proportion shrank to 3.9% in 2019. Newly affected villages as a percentage of all affected villages decreased from 54.3% in 2014 to 23.5% in 2019, as more surveillance data accumulated and overall VL incidence declined. The risk of additional cases in a village increased with increasing cumulative incidence, reaching approximately 90% in villages with 12 cases and 100% in villages with 45 cases, but the vast majority of villages had small cumulative case numbers. The time-to-next-case decreased with increasing case-load. Using a 3-year window (2016–2018), a threshold of seven VL cases at the village level selects 329 villages and yields 23% of cases reported in 2019, while a threshold of three cases selects 1,241 villages and yields 46% of cases reported in 2019. Using a 6-year window increases both effort and yield.ConclusionDecisions on targeting must consider the trade-off between number of villages targeted and yield and will depend upon the operational efficiencies of existing programs and the feasibility of specific ACD approaches. The maintenance of a sensitive, comprehensive VL surveillance system will be crucial to preventing future VL resurgence.

As India moves toward the elimination of visceral leishmaniasis (VL) as a public health problem, comprehensive timely case detection has become increasingly important, in order to reduce the period of infectivity and control outbreaks. During the 2000s, localized research studies suggested that a large percentage of VL cases were never reported in government data. However, assessments conducted from 2013 to 2015 indicated that 85% or more of confirmed cases were eventually captured and reported in surveillance data, albeit with significant delays before diagnosis. Based on methods developed during these assessments, the CARE India team evolved new strategies for active case detection (ACD), applicable at large scale while being sufficiently effective in reducing time to diagnosis. Active case searches are triggered by the report of a confirmed VL case, and comprise two major search mechanisms: 1) case identification based on the index case’s knowledge of other known VL cases and searches in nearby houses (snowballing); and 2) sustained contact over time with a range of private providers, both formal and informal. Simultaneously, house-to-house searches were conducted in 142 villages of 47 blocks during this period. We analyzed data from 5030 VL patients reported in Bihar from January 2018 through July 2019. Of these 3033 were detected passively and 1997 via ACD (15 (0.8%) via house-to-house and 1982 (99.2%) by light touch ACD methods). We constructed multinomial logistic regression models comparing time intervals to diagnosis (30-59, 60-89 and ≥90 days with =90 days compared to the referent of

We have developed an RT-LAMP based protocol for isothermal amplification and subsequent colorimetric detection of the COVID19 infection. This protocol essentially relies upon the RT-LAMP mediated reverse transcription and subsequent amplification into large copies of cDNA products in the first step of the protocol. Subsequently, we have fused a second step where we label the cDNA with a fluorophore based probe to increase the sensitivity and specificity of the results. This step also makes the overall process to be compatible with the paper strip based detection technique where the product from the second step gets captured by the molecule impregnated in the paper strip. This leads to the development of the control and test line which could be detected with a smart phone based app for the prediction and rapid dissemination of the result.

Maps of Bihar showing burdens of identified cases in study districts for January 2012Â -Â June 2013 at (a) district level and (b) block level. (DOCX 996Â kb)

Box-plots of distribution of onset-to-diagnosis waiting times by season of onset. (DOCX 90Â kb)

Single-variable negative-binomial regression models for OD and OT waiting times. (DOCX 23Â kb)

Distribution of continuous variables (age, waiting times and house size) for the eight study districts. (DOCX 15Â kb)

Distribution of socio-economic factors across the eight study districts. (DOCX 30Â kb)

Background Visceral leishmaniasis (VL) is highly prevalent in the Indian state of Bihar and, without proper diagnosis and treatment, is associated with high fatality. However, lack of efficient reporting mechanism had been an impediment in estimating the burden of mortality and its antecedents among symptomatic VL cases. The objectives of the current study were to generate a reliable estimate of symptomatic VL caseload and mortality in Bihar, as well as to identify the epidemiologic and health infrastructure-related predictors of VL mortality. Methodology and Principal Findings Using an elaborate index case tracing method, we attempted to locate all symptomatic VL patients in eight districts of Bihar. Interviews and medical-record-reviews were conducted with cases (or next-of-kin for the dead) meeting the eligibility criteria. The information collected during the interviews included socio-demographic characteristics, onset of disease symptoms, place of diagnosis, pre- and post-diagnosis treatment history, type and duration of drugs received. In total, we analyzed data on 4925 VL patients—59% were male and 68% were less than 30 years old. There were 158 (3.2%) deaths and the incidence rate of mortality was 3.2/100 person-years. In the adjusted Cox-proportional-hazards analysis, treatment at public facility [Adjusted Hazard Ratio (AHR) = 0.61; 95% CI = 0.43–0.86], shorter (≤30 days) diagnostic delay [AHR = 0.62, 95% CI = 0.43–0.92], and treatment completion [AHR = 0.03, 95% CI = 0.02–0.05] emerged as significant negative predictors of mortality. Conclusion Mortality reduction efforts in Bihar should focus on improving access to early diagnosis, quality treatment and treatment-adherence measures, with special emphasis on marginalized communities., Author Summary More than 70% cases of visceral leishmaniasis (VL), a parasitic disease transmitted by sand flies, in India are reported from Bihar, a resource-poor state. In absence of early diagnosis and treatment, the fatality is very high among symptomatic VL cases. However, community-based data on VL mortality is limited in Bihar. Also, little information on the factors associated with mortality among symptomatic VL patients is available from this Indian state. Evidences regarding mortality parameters can prove immensely helpful in designing specific interventions targeted at reduction of such mortality. In the current study, we created a registry of symptomatic VL cases in eight districts of Bihar to estimate the burden of VL mortality and the socio-demographic factors associated with it. The mortality rate among the symptomatic VL patients was found to be fairly high. Shorter interval between onset of symptoms and diagnosis, and completing course of treatment were associated with increased survival. We recommend improving access to early diagnosis and treatment services, especially among the marginalized communities, as a key measure to reduce mortality among symptomatic VL patients in Bihar.