Earthquake precursory detection using diurnal GPS-TEC and kriging interpolation maps: 12 May 2008, Mw7.9 Wenchuan case study.

This study presents an alternative vertical total electron content (VTEC) anomaly detection technique based on diurnal VTEC values. In order to assess the consistency of the results, Mw7.9 Wenchuan earthquake occurred on May 12, 2008 was chosen as case study because several researches were performed on this earthquake event. In this detection technique, the daily mean of VTEC (AVTEC) and standard deviation of diurnal VTEC (SVTEC) were adopted in the analytical process instead of quartile-based technique. The spatial distribution of VTEC was illustrated by AVTEC and SVTEC maps which created from ordinary Kriging interpolation technique. The anomalous day derived from AVTEC and SVTEC was observed on May 9, 2008. The anomalous zone significantly appeared within the earthquake preparation zone in the southeast of the epicenter. The results were corresponding to the previous studies in terms of time and space. Thus, AVTEC, SVTEC and instantaneous ionospheric maps created from ordinary Kriging interpolation technique should be an alternative approach for detecting ionospheric anomaly prior to earthquake occurrence.•Simplified seismo-ionospheric anomaly detection technique•Ionospheric distribution is modelled by ordinary Kriging interpolation maps•The results are consistent with the previous studies in terms of time and space.

Determining suitable machine learning classifier technique for prediction of malaria incidents attributed to climate of Odisha.

This study investigated the influence of climate factors on malaria incidence in the Sundargarh district, Odisha, India. The WEKA machine learning tool was used with two classifier techniques, Multi-Layer Perceptron (MLP) and J48, with three test options, 10-fold cross-validation, percentile split, and supplied test. A comparative analysis was carried out to ascertain the superior model among malaria prediction accuracy techniques in varying climate contexts. The results suggested that J48 had exhibited better skill than MLP with the 10-fold cross-validation method over the percentile split and supplied test options. J48 demonstrated less error (RMSE = 0.6), better kappa = 0.63, and higher accuracy = 0.71), suggesting it as most suitable model. Seasonal variation of temperature and humidity had a better association with malaria incidents than rainfall, and the performance was better during the monsoon and post-monsoon when the incidents are at the peak.

Performance of AnnAGNPS model in predicting runoff and sediment yields in Nan Province, Thailand.

Land use changes such as deforestation and urban development influences the river discharge, soil erosion and sediment yield. It is important to evaluate tools which can be used to assess such impacts on water and sediment yield. Therefore, this study evaluated the Annualized Agricultural Non-Point Source Pollutant (AnnAGNPS) model's performance in simulating runoff and sediment loads in Nan Province, Thailand using seven years of continuous monitoring data. The river discharge and sediment yield data from 2011-2013 were used for calibration, and data from 2014-2017 were used for validation. Several input parameters were computed using methods suggested by other researchers and previous studies. In this study, the runoff curve number, soil erodibility factor (K), and RUSLE-C value were used to accurately simulate runoff and sediment loads. The results indicate that the model satisfactorily simulated runoff and sediment loads (R2 = 0.65 and NSE = 0.53 for runoff volume, and R2 = 0.62 and NSE = 0.60 for sediment yields). Moreover, the model estimated the total sediment yield, which contributed 12,932 hundred tons of material to the Nan River in 2017. The maximum sediment yield was obtained below the catchment (Na Noi sub-district, Na Noi district), which corresponds to areas with high crop densities. Cropland generated the highest soil erosion of all investigated land use (87.52% of total soil erosion). Thus, the AnnAGNPS model has the potential to use for investigating management practices to reduce soil erosion and controlling floods and droughts in Nan Province of Thailand.

Development of a PCR-lateral flow assay for rapid detection of Yersinia pestis, the causative agent of plague.

Plague is a zoonotic disease caused by Yersinia pestis, a Gram-negative, rod shaped coccobacillus, which is primarily found in rodents and can be transmitted to humans through flea bite. The disease has three major clinical forms bubonic (by flea bite), pneumonic (by respiratory droplets) and septicemic plague. Y. pestis is classified as a category 'A' agent by NIAID, USA due to its high mortality and easy person to person dissemination. The conventional diagnostic methods available for Y. pestis show cross-reactivity with other enteropathogenic bacteria making its detection difficult. There is a need to develop sensitive and specific molecular assay for accurate detection of Y. pestis. PCR is well suited molecular biology tool for rapid diagnosis of plague but after completion of thermal cycling steps, it requires additional time to analyze amplified product using agarose gel electrophoresis. In the present study, PCR assay coupled with lateral flow strips has been developed for rapid detection of Y. pestis. Lateral flow strips give an alternative to gel electrophoresis and permit easy and rapid detection of PCR products. The PCR was performed with 5' 6-FAM and biotin tagged primers specific for Y. pestis, targeting yihN gene located on chromosome. The PCR product was analyzed using lateral flow strips which yielded result within 2-3 minutes. The analytical sensitivity of PCR-lateral flow (PCR-LF) assay was 1 pg genomic DNA of Y. pestis and 500 copies of target DNA sequence harboured in a recombinant plasmid. The assay could detect Y. pestis DNA extracted from spiked human blood samples containing ≥104 CFU per mL of bacteria. The assay was found to be specific and did not cross react with other closely related bacterial species. The developed assay was highly specific, sensitive and also did not require agarose gel electrophoresis for post amplification analysis.

Development of a PCR Lateral Flow Assay for Rapid Detection of Bacillus anthracis, the Causative Agent of Anthrax.

Bacillus anthracis, the causative agent of anthrax is one of the most potent listed biological warfare agents. The conventional microbiological methods of its detection are labor intensive and time consuming, whereas molecular assays are fast, sensitive and specific. PCR is one of the most reliable diagnostic tools in molecular biology. The combination of PCR with lateral flow strips can reduce the diagnostic/detection time. It gives an alternative to gel electrophoresis and offers easy and clear interpretation of results. In the present study, a PCR Lateral flow (PCR-LF) assay targeting cya gene present on pXO1 plasmid of B. anthracis has been developed. The forward and reverse primers were tagged with 6-carboxyflourescein (6-FAM) and biotin, respectively, at 5' end. The dual labeled PCR products were detected using lateral flow (LF) strips developed in this study. The PCR-LF assay could detect ≥ 5 pg of genomic DNA and ≥ 500 copies of target DNA harboured in a recombinant plasmid. The assay was able to detect as few as 103 and 10 CFU/mL of B. anthracis Sterne cells spiked in human blood after 6 and 24 h of enrichment, respectively.

Development of a set of three real-time loop-mediated isothermal amplification (LAMP) assays for detection of Bacillus anthracis, the causative agent of anthrax.

Bacillus anthracis, the causative agent of anthrax is a Gram-positive, non-motile, spore forming bacterium. Its spores can persist in soil and water for years and can also be aerosolized. A rapid, sensitive and specific method to detect B. anthracis is important for clinical management and preventing spread of anthrax. Loop-mediated isothermal amplification (LAMP) assay is a rapid technique that amplifies target DNA in isothermal conditions with high sensitivity and specificity. In this study, a LAMP assay set targeting a chromosomal and two plasmid markers was developed. The individual assays of the LAMP set targeting pXO1 plasmid (lef), pXO2 plasmid (capB), and chromosome (BA5345) sequences could detect 10, 250, and 100 fg of genomic DNA and 10, 100, and 50 copies of the DNA targets harboured in recombinant plasmids, respectively. The lef and capB LAMP assays could detect ≥ 1 × 103 CFU per mL of bacteria in spiked human blood samples, while BA5345 LAMP assay could detect ≥ 1 × 104 CFU of bacteria per mL of spiked blood. The amplification was monitored in real-time by turbidimeter, and visual detection was also accomplished under normal and UV light after adding SYBR Green 1 dye on completion of the reaction. The assay set was found to be highly sensitive and did not cross-react with the closely related Bacillus spp. and other bacterial strains used in the study.

Development of a pair of real-time loop mediated isothermal amplification assays for detection of Yersinia pestis, the causative agent of plague.

Yersinia pestis, the causative agent of plague mainly infects rodents, while humans are the accidental host. The conventional diagnostic methods available for Y. pestis exhibit cross-reactivity with other enteropathogenic bacteria which makes its detection difficult. Rapid and reliable point-of-care detection of Y. pestis is essential for timely initiation of medical treatment. In the present study, a pair of loop mediated isothermal amplification (LAMP) assays has been developed for rapid detection of Y. pestis. Two sets of LAMP primers, each containing 6 primers were specifically designed targeting caf1 and 3a genes located on pFra plasmid and chromosome of Y. pestis, respectively. Isothermal amplification was accomplished at 65 °C for 40 min for caf1 target, and at 63 °C for 50 min for 3a choromosomal target. The analytical sensitivity of the assay for the caf1 and 3a targets was found to be 500 fg and 100 fg genomic DNA of Y. pestis, respectively. The caf1 and 3a LAMP assays detected as few as 100 copies of caf1 and 10 copies of 3a gene targets harboured in the respective recombinant plasmids. The amplified products were detected visually under visible and UV light using SYBR Green 1 dye. The assay pair was found to be highly specific as it did not cross-react with closely related and other bacterial species.

Geostatistical predictive modeling for asthma and chronic obstructive pulmonary disease using socioeconomic and environmental determinants.

The spatial distribution of the prevalence of asthma and chronic obstructive pulmonary disease (COPD) remains under the influence of a wide array of environmental, climatic, and socioeconomic determinants. However, a large proportion of these influences remain unexplained. In completion, this study examined the spatial associations between asthma/COPD morbidity and their determinants using ordinary least squares (OLS) and geographically weighted regressions (GWR). Inpatient records collected from the secondary and tertiary care hospitals in Kandy from 2010 to 2014 were considered as the dependent variable. Potential risk factors (explanatory variables) were identified in four distinguished classes: 1) meteorological factors, (2) direct and indirect factors of air pollution, (3) socioeconomic factors, and (4) characteristics of the physical environment. All possible combinations of candidate explanatory variables were evaluated through an exploratory regression. A comparison between the regression models was also explored. The best OLS regression models revealed about 55% of asthma variation and 62% of COPD variation while GWR models yielded 78% and 74% of the variation of asthma and COPD occurrences respectively. Relative humidity, proximity to roads (0-200 m), road density, use of firewood as a source of fuel, and elevation play a vital role in predicting morbidity from asthma and COPD. Both local and global regression models are important in assessing spatial relationships of asthma and COPD. However, the local models exhibit a better prediction capability for assessing non-stationary relationships of asthma and COPD than global models. The geostatistical aspects used in this study may also provide insights for evaluating heterogeneous environmental risk factors in other epidemiological studies across different spatial settings.

Enhanced production and purification of recombinant surface array protein (Sap) for use in detection of Bacillus anthracis.

Surface array protein (Sap) can be an important biomarker for specific detection of Bacillus anthracis, which is released by the bacterium during its growth in culture broth. In the present work, we have cloned and expressed Sap in Escherichia coli. The culture conditions and cultivation media were optimized and used in batch fermentation process for scale up of Sap in soluble form. The recombinant Sap was purified employing affinity chromatography followed by diafiltration. The final yield of purified protein was 20 and 46 mg/l of culture during shake flasks and batch fermentation, respectively. The protein purity and its reactivity were confirmed employing SDS-PAGE and Western blot, respectively. The antibodies raised against purified Sap were evaluated by Western blotting for detection of Sap released by B. anthracis. Our results showed that the Sap could be a novel marker for detection and confirmation of B. anthracis.

A Real-Time Health Monitoring System for Remote Cardiac Patients Using Smartphone and Wearable Sensors.

Online telemedicine systems are useful due to the possibility of timely and efficient healthcare services. These systems are based on advanced wireless and wearable sensor technologies. The rapid growth in technology has remarkably enhanced the scope of remote health monitoring systems. In this paper, a real-time heart monitoring system is developed considering the cost, ease of application, accuracy, and data security. The system is conceptualized to provide an interface between the doctor and the patients for two-way communication. The main purpose of this study is to facilitate the remote cardiac patients in getting latest healthcare services which might not be possible otherwise due to low doctor-to-patient ratio. The developed monitoring system is then evaluated for 40 individuals (aged between 18 and 66 years) using wearable sensors while holding an Android device (i.e., smartphone under supervision of the experts). The performance analysis shows that the proposed system is reliable and helpful due to high speed. The analyses showed that the proposed system is convenient and reliable and ensures data security at low cost. In addition, the developed system is equipped to generate warning messages to the doctor and patient under critical circumstances.

Ecosystem evaluation (1989-2012) of Ramsar wetland Deepor Beel using satellite-derived indices.

The unprecedented urban growth especially in developing countries has laid immense pressure on wetlands, finally threatening their existence altogether. A long-term monitoring of wetland ecosystems is the basis of planning conservation measures for a sustainable development. Deepor Beel, a Ramsar wetland and major storm water basin of the River Brahmaputra in the northeastern region of India, needs particular attention due to its constant degradation over the past decades. A rule-based classification algorithm was developed using Landsat (2011)-derived indices, namely Normalised Difference Water Index (NDWI), Modified Normalised Difference Water Index (MNDWI), Normalised Difference Pond Index (NDPI), Normalised Difference Vegetation Index (NDVI) and field data as ancillary information. Field data, ALOS AVNIR and Google Earth images were used for accuracy assessment. A fuzzy accuracy assessment of the classified data sets showed an overall accuracy of 82 % for MAX criteria and 90 % for RIGHT criteria. The rules were used to classify major wetland cover types during low water season (January) in 1989, 2001 and 2012. The statistical analysis of the classified wetland showed heavy manifestation in aquatic vegetation and other features indicating severe eutrophication over the past 23 years. This degradation was closely related to major contributing anthropogenic factors, such as a railway line construction, growing croplands, waste disposal and illegal human settlements in the wetland catchment. In addition, the landscape development index (LDI) indicated a rapid increase in the impact of the surrounding land use on the wetland from 1989 to 2012. The techniques and results from this study may prove useful for top-down landscape analyses of this and other freshwater wetlands.

Evaluation of a commercial Dengue NS1 enzyme-linked immunosorbent assay for early diagnosis of dengue infection.

PURPOSE: Dengue is one of the most serious mosquito-borne viral infections affecting tropical and subtropical countries in the world. Since there is no immunoprophylactic or specific antiviral therapy available, timely and rapid diagnosis plays a vital role in patient management and implementation of control measures. This paper evaluates a commercially available NS1 antigen capture ELISA vis-a-vis SD bioline Dengue NS1 antigen test for early detection of dengue virus. MATERIALS AND METHODS: To evaluate a commercial NS1 antigen detection kit vis-a-vis SD bioline Dengue NS1 antigen test, a total of 91 clinical samples were tested. Virological investigations with regard to dengue virus, viz. NS1 antigen capture ELISA (Panbio, Australia), SD bioline Dengue NS1 antigen test, RT-PCR and virus isolation were performed. RESULTS: Out of 91 samples, 24 (26%) were positive by NS1 antigen capture ELISA, 15 (16%) by SD bioline Dengue NS1 antigen test and 11(12%) positive by RT-PCR analysis. The RT-PCR-positive samples were further subjected to virus isolation and resulted in three isolates. The results of the Panbio NS1 antigen capture ELISA, SD bioline Dengue NS1 antigen test, RT-PCR and virus isolation were correlated among themselves. CONCLUSIONS: The present study comprehensively established the utility of NS1 antigen ELISA in early diagnosis of dengue infection.

Comparison of a dipstick enzyme-linked immunosorbent assay with commercial assays for detection of Japanese encephalitis virus-specific IgM antibodies.

BACKGROUND: Japanese encephalitis (JE) is a major public health concern in Asia including India. OBJECTIVES: To evaluate an in-house developed dipstick enzyme-linked immunosorbent assay (ELISA) test vis-à-vis two commercial kits for detection of JE virus-specific IgM antibodies. SETTING AND DESIGN: Comparative study carried out in Research and Development centre. MATERIALS AND METHODS: A total of 136 specimens comprising 84 serum and 52 CSF samples were tested by in-house dipstick ELISA, Pan-Bio IgM capture ELISA (Pan-Bio, Australia) and JEV CheX IgM capture ELISA (XCyton, India). RESULTS: The overall agreement among all three tests was found to be 92% with both serum and cerebrospinal fluid (CSF) samples. The sensitivity of the dipstick ELISA was found to be 91% with serum and 89% with CSF samples respectively. The specificity of the dipstick ELISA with reference to both commercial assays was found to be 100% in serum and CSF samples in this study. CONCLUSIONS: The in-house dipstick ELISA with its comparable sensitivity and specificity can be used as a promising test in field conditions since it is simple, rapid and requires no specialized equipment.

Development and evaluation of SYBR Green I-based one-step real-time RT-PCR assay for detection and quantification of Chikungunya virus.

The development of a one-step SYBR Green I-based real-time RT-PCR assay is reported for detection and quantification of Chikungunya virus (CHIKV) in acute-phase patient serum samples by targeting the E1 structural gene. A linear relationship was obtained between the virus concentration and cycle threshold (C(t)) value over a range of 10(7)-0.1PFU/ml. The reported assay was found to be 10-fold more sensitive compared to conventional RT-PCR with a detection limit of 0.1PFU/ml. The feasibility of this reported assay system for clinical diagnosis was validated with 51 suspected acute-phase serum samples of the recent CHIKV epidemic in southern India, 2006. The comparative evaluation with acute-phase patient serum samples revealed the higher sensitivity of real-time RT-PCR assay by picking up six additional samples with low copy number of template. None of the healthy serum samples analyzed in this study showed amplification. The quantification of the viral load in the acute-phase serum samples was also determined employing the standard curve, which varies from 0.1 to 10(7)PFU/ml. These findings demonstrated that the reported assay has the potential usefulness for clinical diagnosis due to simultaneous detection and quantification of Chikungunya virus in acute-phase patient serum samples.

Development and evaluation of SYBR Green I-based one-step real-time RT-PCR assay for detection and quantitation of Japanese encephalitis virus.

One-step SYBR Green I-based real-time RT-PCR assay for rapid detection as well as quantitation of Japanese encephalitis virus (JEV) in acute-phase patient CSF samples by targeting the NS3 gene was developed. The assay developed in this study was found to be more sensitive as compared to conventional RT-PCR. The specificity of the reported assay system was established through melting curve analysis as well as by cross-reactivity studies with related members of Flavivirus. The applicability of Real-time PCR assay for clinical diagnosis was validated with 32 suspected acute-phase CSF samples of Gorakhpur epidemic, India, 2005. The improved sensitivity of real-time RT-PCR was reflected by picking up 10 additional samples with low copy number of template in comparison to conventional RT-PCR. The quantitation of the viral load in acute-phase CSF samples was done using a standard curve obtained by plotting cycle threshold (C(t)) values versus copy numbers of the RNA template. This is the first report on the application of real-time RT-PCR for detection as well as quantitation of JEV from patient CSF samples. These findings demonstrate the potential clinical application of the reported assay as a sensitive diagnostic test for rapid and real-time detection and quantitation of JEV in acute-phase clinical samples.

Rapid and real-time detection of Chikungunya virus by reverse transcription loop-mediated isothermal amplification assay.

The standardization and validation of a one-step, single-tube, accelerated, quantitative reverse transcription (RT) loop-mediated isothermal amplification (RT-LAMP) assay targeting the E1 gene for the rapid and real-time detection of Chikungunya virus (CHIKV) are reported. A linear relationship between the amount of template and time of positivity value over a range of 2 x 10(8) to 2 x 10(2) copies was obtained. The feasibility of CHIKV RT-LAMP for clinical diagnosis was validated with patient serum samples from an ongoing epidemic in Southern India. Optimal assay conditions with zero background were established for the detection of low levels of CHIKV in acute-phase patient serum samples. The comparative evaluation of the RT-LAMP assay with acute-phase patient serum samples demonstrated exceptionally higher sensitivity by correctly identifying 21 additional positive borderline cases that were missed by conventional RT-PCR (P < 0.0001) with a detection limit of 20 copies. The quantification of virus load in patient serum samples was also determined from the standard curve based on their time of positivity and was found to be in the range of 2 x 10(8) to 2 x 10(1) copies. In addition, the field applicability of the RT-LAMP assay was also demonstrated by standardizing SYBR Green I-based RT-LAMP wherein the amplification was carried out in a water bath at 63 degrees C for 60 min, which was followed by monitoring gene amplification with the naked eye through color changes. These findings demonstrated that the RT-LAMP assay is a valuable tool for rapid, real-time detection as well as quantification of CHIKV in acute-phase serum samples without requiring any sophisticated equipment and has potential usefulness for clinical diagnosis and surveillance of CHIKV in developing countries.

East Central South African genotype as the causative agent in reemergence of Chikungunya outbreak in India.

Chikungunya fever is an important arboviral infection prevalent throughout Africa and Southeast Asia. Recently, in 2006, it has reemerged in many parts of India, affecting more than a million persons. A detail serological, virological, and molecular investigation of this unprecedented outbreak was carried out by collecting and studying 540 samples from all the affected regions of India during this epidemic. An in-depth investigation revealed the presence of anti-Chikungunya antibodies in 68% of the samples and genomic RNA in 49% of them. In addition 32 Chikungunya viruses were isolated from 45 representative polymerase chain reaction-positive samples. The nucleotide sequences of partial E1 gene of 25 representative Chikungunya viruses were deciphered. The sequence analysis indicated that all the isolates of this epidemic belonged to the new Indian Ocean island clade of East Central South (ECS) African genotype. This study conclusively proved the genotype shift from Asian to ECS African as the major factor in the reemergence of Chikungunya in an unprecedented outbreak in India after a gap of 32 years.

Development and evaluation of reverse transcription-loop-mediated isothermal amplification assay for rapid and real-time detection of Japanese encephalitis virus.

The standardization and validation of a one-step, single-tube accelerated quantitative reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay is reported for rapid and real-time detection of Japanese encephalitis virus (JEV). The RT-LAMP assay reported in this study is very simple and rapid; the amplification can be obtained in 30 min under isothermal conditions at 63 degrees C by employing a set of six primers targeting the E gene of JEV. The RT-LAMP assay demonstrated exceptionally higher sensitivity compared to that of RT-PCR, with a detection limit of 0.1 PFU. The specificities of the selected primer sets were established by cross-reactivity studies with other closely related members of the JEV serocomplex as well as by evaluation of healthy human volunteers. The comparative evaluation of the RT-LAMP assay for clinical diagnosis with a limited number of patient cerebrospinal fluid samples revealed 85% concordance with conventional RT-PCR, with a sensitivity and a specificity of 100% and 86%, respectively. The concentration of virus in most of the clinical samples was 10(2) to 10(5) PFU/ml, as determined from the standard curve based on the time of positivity in the samples. In addition, the monitoring of gene amplification can also be visualized with the naked eye by using SYBR green I fluorescent dye. Thus, due to easy operation without a requirement of sophisticated equipment and skilled personnel, the RT-LAMP assay reported here is a valuable tool for the rapid and real-time detection of JEV not only by well-equipped laboratories but also by peripheral diagnostic laboratories with limited financial resources in developing countries.