Molecular Evolution of GII.P31/GII.4_Sydney_2012 Norovirus over a Decade in a Clinic in Japan.

Norovirus (NoV) genogroup II, polymerase type P31, capsid genotype 4, Sydney_2012 variant (GII.P31/GII.4_Sydney_2012) has been circulating at high levels for over a decade, raising the question of whether this strain is undergoing molecular alterations without demonstrating a substantial phylogenetic difference. Here, we applied next-generation sequencing to learn more about the genetic diversity of 14 GII.P31/GII.4_Sydney_2012 strains that caused epidemics in a specific region of Japan, with 12 from Kyoto and 2 from Shizuoka, between 2012 and 2022, with an emphasis on amino acid (aa) differences in all three ORFs. We found numerous notable aa alterations in antigenic locations in the capsid region (ORF2) as well as in other ORFs. In all three ORFs, earlier strains (2013-2016) remained phylogenetically distinct from later strains (2019-2022). This research is expected to shed light on the evolutionary properties of dominating GII.P31/GII.4_Sydney_2012 strains, which could provide useful information for viral diarrhea prevention and treatment.

Role of booster vaccines and hybrid immunity against severe COVID-19 outcomes during BA.5 omicron predominance in Thailand.

Owing to both vaccine- and infection-induced immunity, the COVID-19 seroprevalence is ~90% in most countries. It is important to examine the protective role of booster vaccines and hybrid immunity in the COVID-endemic state. Utilizing a hospital information system for COVID-19, we conducted a cohort study by linking laboratory-confirmed COVID-19 case data to the national immunization records during the BA.5 omicron predominant period (1 August-31 December 2022) in Chiang Mai, Thailand. Out of 63,009 adults with COVID-19 included in the study, there were 125 (0.2%) severe COVID outcomes and 6.4% had a previous omicron infection. Protection against severe COVID-19 was highest among those with at least one booster vaccine (63%; aHR 0.37 [95%CI 0.19-0.73]) as compared to those without prior vaccination or natural infection. Hybrid immunity offered better protection (35%; aHR 0.65 [95%CI 0.09-4.73) than primary vaccine series alone or previous infection alone. Evaluating risk by age group, those aged 70 years or more had nearly 40 times (aHR 39.58 [95%CI 18.92-82.79]) the risk of severe-COVID-19 as compared to the 18-39-year age group. While booster vaccines remain the most effective way of protecting against severe COVID-19, particularly in the elderly, hybrid immunity may offer additional benefit.

Heterologous booster vaccines reduce severity and mortality in COVID-19 during BA.2 and BA.4/BA.5 omicron predominance in Thailand.

BACKGROUND: The COVID-19 pandemic has evolved quickly, with variants of concern resulting in the need to offer booster vaccinations. Unfortunately, the booster uptake has been slow and vaccine response has shown to wane over time. Therefore, it's critical to evaluate the role of vaccinations on outcomes with newer sub-lineages of omicron. METHODS: Utilising a Hospital Information System established in Chiang Mai, Thailand, we conducted a cohort study by linking patient-level data of laboratory-confirmed COVID-19 cases to the national immunization records, during BA.2 and BA.4/BA.5 predominance. RESULTS: In adjusted cox-proportional hazard models, BA.4/BA.5 was not associated with more severe COVID-19 outcomes or deaths as compared to BA.2. Risk of severe outcomes and deaths were significantly reduced with third (87% and 95%) and fourth (88% and 95%) dose vaccination, while events were not observed with a fifth dose. Across the regimens, vaccination within 14-90 days prior showed the highest level of protection. All the vaccine types used for boosting in Thailand offered similar protection against severe COVID-19. CONCLUSIONS: Boosters provide high level of protection against severe COVID-19 outcomes and deaths with newer omicron sub-lineages. Booster campaigns should focus on improving coverage utilising all available vaccines to ensure optimal protection.

Waning vaccine response to severe COVID-19 outcomes during omicron predominance in Thailand.

BACKGROUND: The COVID-19 pandemic has evolved quickly, with different variants of concern resulting in the need to offer continued protection through booster vaccinations. The duration of enhanced protection with booster doses against severe COVID-19 is still unclear. Understanding this is critical to recommendations on the frequency of future booster doses. METHODS: Utilising a Hospital Information System for COVID-19 established in Chiang Mai, Thailand, we conducted a cohort study by linking patient-level data of laboratory-confirmed COVID-19 cases to the national immunization records, during the omicron predominant period (1 February- 31 July 2022). RESULTS: Out of 261,103 adults with COVID-19 included in the study, there were 333 (0.13%) severe COVID-19 cases and 190 (0.07%) deaths. Protection against severe COVID-19 was highest with boosters received >14-60 days prior to positive test (93%) and persisted at >60-120 days (91%) but started to wane at >120-180 days (77%) and further at >180 days (68%). The rate of waning differed with age. Those ≥70 years showed faster waning of booster vaccine responses as compared to those aged 18-49 years, who retained good responses up to 180 days. Equivalent risk reduction against severe COVID-19 was seen with all the vaccine types used as boosters in Thailand. CONCLUSIONS: Booster doses provided high levels of protection against severe COVID-19 with omicron, up to 4 months. Repeat boosters will be required to continue protection beyond 4 months, particularly in the elderly. mRNA and viral vector vaccines can be used flexibly to improve booster coverage.

Molecular Epidemiology of Classic, MLB, and VA Astroviruses in Children with Acute Gastroenteritis, 2014-2021: Emergence of MLB3 Strain in Japan.

Human astroviruses (HAstVs) are important causative pathogens of acute gastroenteritis (AGE) in children worldwide. MLB and VA HAstVs, which are genetically distinct from the previously known classic HAstVs, have been detected since 2008. To investigate the role of HAstVs in AGE, we conducted molecular detection and characterization of HAstVs circulating in children with AGE in Japan from 2014 to 2021. Out of 2,841 stool samples, HAstVs were detected in 130 (4.6%). MLB1 was the predominant genotype detected (45.4%), followed by HAstV1 (39.2%), MLB2 (7.4%), VA2 (3.1%), HAstV3 (2.3%), HAstV4, HAstV5, and MLB3 (0.8% each). The results demonstrated that HAstV infection in pediatric patients in Japan was dominated by the two major genotypes MLB1 and HAstV1, with a small proportion of other genotypes. The overall infection rates of MLB and VA HAstVs were higher than those of classic HAstVs. The HAstV1 strains detected in this study belonged solely to lineage 1a. The rare MLB3 genotype was detected for the first time in Japan. All three HAstV3 strains belonged to lineage 3c based on the ORF2 nucleotide sequence and were shown to be recombinant strains. IMPORTANCE HAstVs are one of the pathogens of viral AGE and are considered the third most common viral agents of AGE after rotavirus and norovirus. HAstVs are also suspected to be the causative agents of encephalitis or meningitis in immunocompromised patients and elderly persons. However, little is known about the epidemiology of HAstVs in Japan, especially that of MLBs and VA HAstVs. This study demonstrated epidemiological features and molecular characterization of human astroviruses encompassing a 7-year study period in Japan. This study highlights the genetic diversity of HAstV circulating in pediatric patients with acute AGE in Japan.

Stigmatizing and discriminatory attitudes toward people living with HIV/AIDS (PLWHA) among general adult population: the results from the 6th Thai National Health Examination Survey (NHES VI).

Background: Thailand has an ongoing action plan to reduce human immunodeficiency virus (HIV) discrimination and stigma. We aimed to monitor the level of stigmatizing and discriminatory attitudes toward people living with HIV/AIDS (PLWHA) among the general adult population and to investigate its related factors. Methods: This study was based on data from the 6th Thai National Health Examination Survey, a large-scale country-wide survey in 2019-2020. We used a multistage sampling technique and included 11 843 adults aged 20 to 59. We collected data through face-to-face interviews which included six items related to HIV stigma domains. We weighted all analyses to account for the probability of sampling the Thai population aged 20 to 59 years. Results: We found that anticipated stigma had the highest percentage of negative stigmatizing attitude responses (78.5%), followed by perceived stigma (66.6%), fear of HIV infection (54.4%), and social judgment (28.2%). Regarding the UNAIDS global indicator for discriminatory attitude, 48.6% of respondents had negative perceptions to questions about experienced stigma or discrimination. Multiple logistic regression showed that factors associated with discriminatory attitudes toward PLWHA were being aged 20-39 (adjusted odds ratio (aOR) = 1.32, 95% confidence interval (CI) = 1.18-1.47) or 50-59 (aOR = 1.23, 95% CI = 1.09-1.40) compared to being aged 40-49, being Muslim compared to Buddhist (aOR = 1.73, 95% CI = 1.46-2.06), being married compared to being single (aOR = 1.15, 95% CI = 1.04-1.28), holding certificate degree or higher compared to not studying or studying at a primary level (aOR = 0.81, 95% CI = 0.68-0.97), living in the Northeast (aOR = 1.27, 95% CI = 1.12-1.45) and Bangkok (aOR = 1.30, 95% CI = 1.12-1.51) compared to living in the North, having no HIV/AIDS infected relative or acquaintance compared to having an HIV/AIDS infected relative or acquaintance (aOR = 1.56, 95% CI = 1.41-1.73), and not obtaining an HIV test compared to obtaining it (aOR = 1.10, 95% CI = 1.02-1.19). Conclusions: We found that HIV stigmatizing and discriminatory attitudes toward PLWHA decreased, but remained concerning among Thai adult people. A public education and awareness campaign, as well as an intervention to reduce HIV-related stigma and discrimination in the country's health care facilities, must still be maintained.

Reduction in severity and mortality in COVID-19 patients owing to heterologous third and fourth-dose vaccines during the periods of delta and omicron predominance in Thailand.

OBJECTIVES: The COVID-19 pandemic has evolved quickly, with different variants of concern resulting in the need for countries to offer booster vaccinations. Although studies have assessed homologous schedules in detail, the effectiveness of heterologous booster vaccine schedules against severity and mortality with newer variants remains to be explored fully. METHODS: Utilizing a Hospital Information System for COVID-19 established in Chiang Mai, Thailand, we conducted a cohort study by linking patient-level data on laboratory-confirmed COVID-19 cases to the national immunization records, during delta-predominant and omicron-predominant periods. RESULTS: Compared to omicron, COVID-19 cases during the delta period were 10 times more likely to have severe outcomes and in-hospital deaths. During omicron, a third vaccine dose had an 89% reduced risk of both severe COVID-19 and death. The third dose received 14-90 days before the date of the positive test showed the highest protection (93%). Severe outcomes were not observed with the third dose during delta, and the fourth dose during the omicron period. All the vaccine types used for boosting in Thailand offered similar protection against severe COVID-19. CONCLUSION: Booster doses provided a very high level of protection against severe COVID-19 outcomes and deaths. Booster campaigns should focus on improving coverage by utilizing all available vaccines to ensure optimal protection.

Effectiveness of heterologous third and fourth dose COVID-19 vaccine schedules for SARS-CoV-2 infection during delta and omicron predominance in Thailand: a test-negative, case-control study.

Background: The Coronavirus disease 2019 (COVID-19) pandemic has evolved quickly, with numerous waves of different variants of concern resulting in the need for countries to offer continued protection through booster vaccination. To ensure adequate vaccination coverage, Thailand has proactively adopted heterologous vaccination schedules. While randomised controlled trials have assessed homologous schedules in detail, limited data has been reported for heterologous vaccine effectiveness (VE). Methods: Utilising a unique active surveillance network established in Chiang Mai, Northern Thailand, we conducted a test-negative case control study to assess the VE of heterologous third and fourth dose schedules against SARS-CoV-2 infection among suspect-cases during Oct 1-Dec 31, 2021 (delta-predominant) and Feb 1-Apr 10, 2022 (omicron-predominant) periods. Findings: After a third dose, effectiveness against delta infection was high (adjusted VE 97%, 95% CI 94-99%) in comparison to moderate protection against omicron (adjusted VE 31%, 95% CI 26-36%). Good protection was observed after a fourth dose (adjusted VE 75%, 95% CI 71-80%). VE was consistent across age groups for both delta and omicron infection. The VE of third or fourth doses against omicron infection were equivalent for the three main vaccines used for boosting in Thailand, suggesting coverage, rather than vaccine type is a much stronger predictor of protection. Interpretation: Appropriately timed booster doses have a high probability of preventing COVID-19 infection with both delta and omicron variants. Our evidence supports the need for ongoing national efforts to increase population coverage of booster doses. Funding: This research was supported by the National Research Council of Thailand (NRCT) under The Smart Emergency Care Services Integration (SECSI) project to Faculty of Public Health Chiang Mai University.

Genotypic Distribution and the Epidemiology of Multidrug Resistant Tuberculosis in Upper Northern Thailand.

The epidemiology and genotypes of multidrug-resistant tuberculosis (MDR-TB), a global public health threat, remain limited. The genotypic distribution and factors associated with MDR-TB in upper northern Thailand between 2015 and 2019 were investigated. The DNA sequencing of rpoB, katG, and inhA promoter of 51 multidrug-resistant Mycobacterium tuberculosis isolates revealed nine patterns of the rpoB gene mutation distributed in seven provinces. The S531L mutation was the most common mutation in all provinces. The rpoB mutation in Chiang Rai, Chiang Mai, and Lampang was highly diverse compared to other areas. Here, the mutation profiles that have yet to be reported in northern Thailand (H526P, Q513P, and H526C) were detected in Chiang Rai province. The S315T katG mutation was the most common genotype associated with INH resistance, especially in Chiang Mai and Lampang. Further analysis of data from 110 TB patients (42 MDR-TB and 68 drug-susceptible TB) revealed that <60 years of age was a significant factor associated with MDR-TB (OR = 0.316, 95% CI 0.128−0.784, p = 0.011) and ≥60 years of age was a significant factor associated with the S315T katG-mutation (OR = 8.867, 95% CI 0.981−80.177, p = 0.047). This study highlighted the necessity for continuous surveillance and risk factor monitoring for effective control of MDR-TB.

Changing distribution of rotavirus A genotypes circulating in Japanese children with acute gastroenteritis in outpatient clinic, 2014-2020.

BACKGROUND: Rotavirus A (RVA) is a major cause of severe acute gastroenteritis (AGE) in infants and children worldwide. In Japan, two kinds of rotavirus vaccines have been introduced as voluntary vaccines in 2011 and 2012, respectively, and launched into the national vaccine program in October 2020. METHODS: In this study, we investigated prevalence of RVA and their molecular characterization in the stool samples collected from infants and children with AGE who visited one outpatient clinic in Japan, from July 2014 to June 2020, during voluntary vaccination with two kinds of rotavirus vaccines. RESULTS: The RVA detection rates decreased from 44.7 % in 2014-2015 to 35.4 % in 2018-2019, whereas in 2019-2020 the numbers of samples collected were dramatically decreased and none of RVA was detected. During this study period, rotavirus vaccination rates in this area increased from 32.4 % to 62.2 %. Distribution of RVA VP7 (G), VP4 (P), and VP6 (I) genotypes in this area had changed year by year; the major genotype combinations were G1P[8]I1 and G1P[8]I2 in 2014-2015, G2P[4]I2 and G9P[8]I1 in 2015-2016, G1P[8]I1 and G8P[8]I2 in 2017-2018, and G8P[8]I2 in 2018-2019. Phylogenetic analysis demonstrated that VP7 nucleotide sequences of G1 were genetically diverse compared with those of other G genotypes in this study. Meanwhile, predominance of unusual G2P[8]I1, G2P[8]I2 and mixed P genotypes were observed only in 2016-2017, but did not carry on in 2017-2019. The equine-like G3 was detected only in 2016-2017. CONCLUSIONS: The results revealed diversity of RVA genotypes and the genotype combinations have changed year by year in Japan, during the study period of 2016-2020.

An increasing trend of human sapovirus infection in Japan, 2009 to 2019: An emerging public health concern.

BACKGROUND: Human sapovirus (SaV) is an important etiologic agent of childhood diarrhea. This study aims to investigate the burden of SaV infection in childhood diarrhea in Japan from 2009-2019, to understand the changes in SaV infection after the introduction of rotavirus (RV) vaccination in Japan in 2011. METHODS: Stool samples were collected from children aged ≤ 12 years old with acute gastroenteritis (AGE) who visited outpatient clinics of six prefectures in Japan. The viral RNA was detected by RT-PCR and genogroups and genotypes were determined through sequence-based analysis. RESULTS: Among 5697 stool samples, 318 (5.6%) samples remained SaV-positives showing the highest prevalence in June and 12-24 month aged children. The most predominant genotype was GI.1 (56.8%), followed by GI.2 (19.2%), GII.1 (10.8%), GIV.1 (9.4%), GI.3 (1.7%), GII.2 (1.4%), GII.3 and GII.5 (0.3%). Importantly, an increasing trend (P = 0.016) of SaV infection was observed during this period. In particular, SaV-detection rate was increased significantly (P = 0.033) from 4.3% in pre-rotavirus (RV)-vaccination era to 6.1% in post-RV-vaccination era. We provided evidence that this increase in SaV infection was mainly attributed by coinfections. CONCLUSIONS: The upward trend of SaV infection, particularly after the introduction of RV-vaccination, is an emerging concern. Attention should be paid to control this upward trend of SaV infection to ensure maximum benefits of implementation of RV vaccines towards reducing overall childhood diarrhea worldwide.

Diversity of human sapovirus genotypes detected in Japanese pediatric patients with acute gastroenteritis, 2014-2017.

Sapovirus (SaV) is one of the pathogens related to acute gastroenteritis (AGE) in adults and children worldwide. This study reported the diversity of SaV genotypes in children with AGE in Japan from July 2014 to June 2017. Of a total of 2259 stool samples tested by using reverse transcription-PCR method and further analyzed by nucleotide sequencing, 114 (5.0%) were positive for SaV and GI.1 (83.3%) was the most predominant genotype, followed by GII.1, GIV.1, GI.2, GI.3, and GII.3 genotypes. Monthly distribution analysis demonstrated two epidemic peaks from July to December 2015 and February to May 2017. However, no detection peak was observed in 2014 and 2016. Phylogenetic analysis of the complete VP1 nucleotide sequences of these GI.1 strains revealed two major clusters of GI.1 and each of which contained GI.1 strains of both 2015 and 2017. This study suggests that the continuous surveillance of SaV is needed to monitor high genetic diversity in Japanese children with AGE.

Genetic diversity of Parechovirus A in infants and children with acute gastroenteritis in Japan during 2016-2018.

Parechovirus A (PeV-A), previously known as human parechovirus, is a common pathogen in children that can cause respiratory and gastrointestinal diseases as well as severe neurological disease. Take advantage of our previous findings on the genetic diversity of PeV-A circulating in Japanese children with acute gastroenteritis (AGE), this study was conducted to investigate the genetic diversity of PeV-A isolated from children with AGE in Japan as well as their clinical symptoms. Of 1070 stool samples collected from Japanese infants and children with AGE during the 2-year period from July 2016 to June 2018, 76 were positive for PeV-A by multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) and were subjected to genotyping based on viral protein 1 (VP1) sequences. Five different PeV-A genotypes including PeV-A1B, -A2, -A3, -A4, and -A6 were detected with predominant of PeV-A1 clade B genotype. This study revealed a high genetic diversity of PeV-A circulating in Japanese infants and children with AGE and the PeV-A2, a rare genotype, was detected for the first time in Japan in patients with AGE. The clinical symptoms observed in these patients included diarrhea, vomiting, fever, cough, rhinorrhea, and dehydration.

Genetic diversity of norovirus genogroup I, II, IV and sapovirus in environmental water in Thailand.

BACKGROUND: Enteric caliciviruses, including noroviruses (NoVs) and sapoviruses (SaVs) are the most significant pathogens associated with waterborne and foodborne outbreaks of nonbacterial acute gastroenteritis in humans worldwide. METHODS: In this study, 126 environmental water samples collected from 6 different sources in Chiang Mai, Thailand from November 2016 to July 2018 were examined for the presence of genogroups I, II, IV (GI, GII, GIV) NoVs and SaVs by using RT-nested PCR assays, genome sequencing, and phylogenetic analysis, RESULTS: Forty out of 126 (31.7%) water samples were positive for one or more caliciviruses throughout the years of study with high prevalence in winter. Among 126 tested specimens, 34 (27.0%), 30 (23.8%), 3 (2.4%), and 2 (1.6%) were positive for NoV GI, GII, GIV, and SaV, respectively. For NoV GI, 6 different genotypes were identified with the most predominant of GI.1 genotype (17 strains). In addition, 6 different genotypes of GII were detected with high prevalence of GII.17 (12 strains) and GII.2 (11 strains). It was interesting to note that our study reported the detection of NoV GIV for the first time in water samples in Thailand, and all were GIV.1 genotype. For SaV detection, only 2 water samples were positive for SaV GI. CONCLUSIONS: The data revealed heterogeneity and highly dynamic distribution of NoV GI, GII, GIV, and SaV in environmental water in Chiang Mai, Thailand, during the study period of 2016-2018.

Distribution of rotavirus genotypes in Japan from 2015 to 2018: Diversity in genotypes before and after introduction of rotavirus vaccines.

BACKGROUND: Diversity in group A rotavirus (RVA) strains after introduction of RV-vaccines remains an emerging concern worldwide. In this study, we investigated the prevalence and distribution of RVA genotypes in Japanese children with acute gastroenteritis (AGE) from 2015 to 2018. In addition, a comparison of the genotypes in pre-vaccination (2006-2012) and post-vaccination (2012-2018) periods was conducted to understand the impact of these vaccines on genotype distribution. METHODS: Fecal samples were collected regularly from outpatient clinics in six localities: Hokkaido, Tokyo, Shizuoka, Osaka, Kyoto, and Saga. RVA were screened and genotyped by RT-PCR and sequence-based genotyping. RESULTS: During the period 2015-2018, RVA was detected in 307 (19.7%) samples out of 1557 specimens: 29.9% (95% CI: 25.8% to 34.3%), 17.9% (95% CI: 14.7% to 21.5%), and 13% (95% CI: 10.3% to 16.0%) were detected RVA-positive in 2015-2016, 2016-2017 and 2017-2018, respectively. The average detection of RVA in pre-vaccination (2006-2012) and post-vaccination (2012-2018) era remained almost similar (18%-20%). The G2P[4]I2 (52.1%, 95% CI: 43.5%-60.6%) remained the most common genotype in 2015-2016, whereas G8P[8]I2 (55.9%, 95% CI: 45.2%-66.2%) dominated in 2016-2017. In 2017-2018, G9P[8]I2 (42.0%, 95% CI: 30.5%-53.9%) prevailed, followed by G9P[8]I1 (23.0%, 95% CI: 14.0%-34.2%). The detection rate of some common genotypes of pre-vaccination era like G1P[8] and G3P[8] has been reduced after introduction of RV-vaccine, whereas genotypes that were sporadic before the introduction of vaccines like G2P[4], G2P[8], G9P[8] and G8P[8] were emerged/reemerged in post-vaccination period. CONCLUSIONS: Our study presented the diversity in circulating RVA genotypes in Japan before and after introduction of RV-vaccines. Sudden emergence of DS-1-like (I2) unusual strains in post-vaccination era remains alarming. Continuous monitoring of RVA genotypes is therefore indispensable to refine future vaccine strategy.

Molecular detection of enteric viruses in the stool samples of children without diarrhea in Bangladesh.

A number of molecular epidemiological studies reported the detection of enteric viruses in asymptomatic children. The role of these viruses in an asymptomatic infection remains unclear. This study investigated the enteric viruses in the stool samples collected from children without diarrhea. Stool samples were collected during June to October 2016, from 227 children who lived in Matlab, Bangladesh. Seventeen enteric viruses, including rotavirus A, B, and C (RVA, RVB, and RVC), norovirus GI (NoV GI), norovirus GII (NoV GII), sapovirus (SaV), adenovirus (AdV), human astrovirus (HAstV), Aichivirus (AiV), human parechovirus (HPeV), enterovirus (EV), human bocavirus (HBoV), Saffold virus (SAFV), human cosavirus (HCoSV), bufavirus (BufV), salivirus (SalV), and rosavirus (RoV), were investigated by RT-PCR method. One hundred and eighty-two (80.2%; 182/227) samples were positive for some of these viruses, and 19.8% (45/227) were negative. Among the positive samples, 46.7% (85/182) were a single infection, and 53.3% (97/182) were coinfection with multiple viruses. The HCoSV was the most prevalent virus (41.4%), followed by EV (32.2%), NoV GII (25.6%), HPeV (8.8%), RVA (6.2%), AdV (5.7%), AiV (5.3%), SAFV (4.4%), and SaV (2.6%). Each of NoV GI, HAstV, HBoV, and BufV was detected at 0.4%. However, RVB, RVC, SalV, and RoV were not detected in this study. Phylogenetic analysis showed that diverse HCoSV species and genotypes were circulating in Bangladesh, and four strains of species A are proposed to be new genotypes. The data indicated that non-diarrheal Bangladeshi children were asymptomatically infected with wide varieties of enteric viruses.

Evaluation of Immunochromatographic Tests for Detection of a Wide Variety of Group A Rotavirus Genotypes and Adenovirus.

BACKGROUND: Viral gastroenteritis is one of the most common illnesses in humans worldwide, and different viral agents have been shown to be associated with the disease. Among these, rotaviruses and adenoviruses are the responsible causative agents of acute gastroenteritis and causing numerous outbreaks. Therefore, a simple and rapid diagnostic tool, such as an immunochromatographic (IC) test, is required for rapid diagnosis, especially during an outbreak of these pathogens. METHODS: The efficiency of two commercial IC kits were evaluated for simultaneous detections of rotavirus and adenovirus in clinical stool specimens by a single test kit. RESULTS: The data demonstrated that both IC test kits could detect either adenovirus or rotavirus positive alone, as well as mixed infections of both viruses in a single stool specimen. In addition, a wide variety of rotavirus genotypes, including G1-P[8]-I1, G2-P[4]-I2, G3-P[8]-I2, G8-P[8]-I2, and G9-P[8]-I1 could be detected by both IC kits. The detection limit of the kits for the detection of rotavirus and adenovirus were comparable to those of real-time PCR at 105 copies/mL. CONCLUSIONS: These two IC test kits could be used as an alternative choice for rapid screening of rotavirus and adenovirus in the stool specimens, especially during the seasonal outbreak of acute gastroenteritis.

Diversity of human parechovirus in infants and children with acute gastroenteritis in Japan during 2014-2016.

A total of 972 stool samples were collected from infants and children with acute gastroenteritis (AGE) in pediatric clinics encompassing six localities (Hokkaido, Tokyo, Shizuoka, Kyoto, Osaka, and Saga) in Japan during the 2-year period from July 2014 to June 2016. Sixty six of the samples (6.8%) were found to be positive for human parechovirus (HPeV) by multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) and subjected to genotyping based on viral protein 1 (VP1) sequences. Four different HPeV genotypes consisting of HPeV1, -3, -4 and -6 were detected, with HPeV1 clade B being predominant and followed by HPeV3 and -6. The first-time presence of HPeV1 clade A in Japan and rare HPeV4 were noted. This study provides up-to-date information on the genetic diversity of HPeV circulating in Japanese infants and children with AGE.

Alarming Situation of Spreading Enteric Viruses Through Sewage Water in Dhaka City: Molecular Epidemiological Evidences.

Global burden of acute viral gastroenteritis remains high, particularly in developing countries including Bangladesh. Sewage water (SW) is an important node to monitor enteric pathogens both in the environment and among the population. Analysis of SW in Dhaka city deems crucially important because a large number of urban-city dwellers live in Dhaka city, the capital of Bangladesh, under a constant threat of precarious sewerage system. In this study, we collected raw SW from five locations of Dhaka city every month from June 2016 to May 2017. It was concentrated with polyethylene glycol (PEG) and investigated for three major enteric viruses, rotavirus A (RVA), norovirus GII (NoV GII) and adenovirus (AdV) using polymerase chain reaction (PCR). Most of these SW samples collected from both hospitals and non-hospital areas yielded enteric viruses: 76% samples were positive for AdV, followed by 53% NoV GII and 38% RVA. Viral load was determined as much as 1 × 107 copies/ml for RVA and 3.5 × 103 copies/ml for NoV GII. Importantly, NoV GII and AdV that can affect people of all ages were predominated during monsoon also when SW overflows and spreads over a wide and crowded area. Genotypes G1, G2, G3, G8, and G9 for RVA, GII.4 for NoV, and type 41 for AdV were detected representing the current profile of circulating genotypes in the population. This study provides the first evidence of distribution of major diarrheal viruses in SW in Dhaka city which is alarming showing grave risk of impending outbreaks through exposure.

Evaluation of Immunochromatographic Test for Dual Detection of Noroviruses and Group A Rotaviruses in Stool Samples.

BACKGROUND: Viral enteric infections, such as noroviruses and rotaviruses are considered as the major causes of acute gastroenteritis in young children and elderly people all over the world. These viruses are responsible for numerous outbreaks of nonbacterial gastroenteritis in several settings such as hospitals, day care centers, nursing homes, and restaurants. Therefore, a rapid and sensitive diagnostic tool for virus detections could be helpful for disease control. METHODS: A new immunochromatographic test for the dual detection of noroviruses and group A rotaviruses was evaluated by using specimens that were known to be positive for noroviruses and rotaviruses, as well as other diarrheal viruses. RESULTS: The sensitivity of detections for noroviruses and group A rotaviruses were 89.5% and 100%, respectively, while the specificity was 100% for both viruses. This immunochromatographic test was reactive to several norovirus and group A rotavirus genotypes (NoV GII.2, GII.3, GII.4, GII.6, GII.7, GII.17 and RVA G1P[8], G2P[4], G3P[8], G8P[8], G9P[8]). CONCLUSIONS: The speed and ease of use of the immunochromatographic test makes this kit particularly attractive as an alternative method for the detections of noroviruses and group A rotaviruses in the primary care unit.