Evaluation of different genomic regions of rotavirus B and rotavirus C for development of real-time RT‒PCR assays.

BACKGROUND: The causative agents of diarrhea, rotavirus B (RVB) and rotavirus C (RVC) are common in adults and patients of all age groups, respectively. Due to the Rotavirus A (RVA) vaccination program, a significant decrease in the number of gastroenteritis cases has been observed globally. The replacement of RVA infections with RVB, RVC, or other related serogroups is suspected due to the possibility of reducing natural selective constraints due to RVA infections. The data available on RVB and RVC incidence are scant due to the lack of cheap and rapid commercial diagnostic assays and the focus on RVA infections. The present study aimed to develop real-time RT‒PCR assays using the data from all genomic RNA segments of human RVB and RVC strains available in the Gene Bank. RESULTS: Among the 11 gene segments, NSP3 and NSP5 of RVB and the VP6 gene of RVC were found to be suitable for real-time RT‒PCR (qRT‒PCR) assays. Fecal specimens collected from diarrheal patients were tested simultaneously for the presence of RVB (n = 192) and RVC (n = 188) using the respective conventional RT‒PCR and newly developed qRT‒PCR assays. All RVB- and RVC-positive specimens were reactive in their respective qRT‒PCR assays and had Ct values ranging between 23.69 and 41.97 and 11.49 and 36.05, respectively. All known positive and negative specimens for other viral agents were nonreactive, and comparative analysis showed 100% concordance with conventional RT‒PCR assays. CONCLUSIONS: The suitability of the NSP5 gene of RVB and the VP6 gene of RVC was verified via qRT‒PCR assays, which showed 100% sensitivity and specificity. The rapid qRT‒PCR assays developed will be useful diagnostic tools, especially during diarrheal outbreaks for testing non-RVA rotaviral agents and reducing the unnecessary use of antibiotics.

Comparing single versus multiple virus detection in pediatric acute gastroenteritis postimplementation of routine multiplex RT-PCR diagnostic testing.

Utilizing multiplex real time polymerase chain reaction (RT-PCR) for rapid diagnosis of gastroenteritis, enables simultaneous detection of multiple pathogens. A comparative analysis of disease characteristics was conducted between cases with single and multiple viruses. Rotavirus vaccine was introduced in 2010, reaching a 70% coverage in 2 years. All rectal swabs collected from diarrheic children (<5 years) between December 2017 and March 2022 were included. Detection of the same viruses within 2 months was considered a single episode. Episodes with positive stool bacterial PCR were excluded. A total of 5879 samples were collected, revealing 86.9% (1509) with single virus detection and 13.1% (227) with multiple viruses. The most frequent combination was rotavirus and norovirus (27.8%), these infections followed a winter-spring seasonality akin to rotavirus. Children with multivirus infections exhibited higher immunodeficiency (OR 2.06) rates, but lower food allergy (OR 0.45) and prematurity rates (OR 0.55) compared to single infections. Greater disease severity, evaluated by the Vesikari score, was observed in multivirus episodes (p < 0.001, OR 1.12). Multivirus infections accounted for 13.1% of symptomatic cases in hospitalized young children. Despite vaccination efforts, rotavirus remained prominent, frequently in co-infections with norovirus. Overall, multivirus infections were linked to more severe diseases than single virus cases.

No sign of Rotavirus co-infection in COVID-19 patients with gastrointestinal symptoms.

Background and aims: The main goal of the present study is to investigate the incidence of Rotavirus co-infection in COVID-19 patients. Methods and Results: Fecal samples of COVID-19 patients with gastrointestinal symptoms which had positive PCR- were collected from Abadan's hospital, Iran during the period December 2020 to January 2021. Samples were analyzed by RT-PCR to determine the presence of Rotavirus. Finally, the total samples size of 37 were included in this study. The mean age of patients was 48.22 years. Abdominal pain alone was detected in 48.65% of the patients. At least one gastrointestinal symptom was detected in all of the patients. Diarrhea and fever were seen in 13.51% and 59.46% of patients, respectively. Nausea and vomiting were seen in 5.41% of the patients. RT-PCR showed no infection of Rotavirus among the patients. Conclusion: Gastrointestinal symptoms related to COVID-19 are common. More studies is need among these patients groups for investigate co-infection with other fecal viral shedding carries, due to a worse prognosis and its association with disease severity.

Application of the cell-based RT-qPCR assay (C-QPA) for potency detection of the novel trivalent rotavirus vaccine in China.

BACKGROUND: Because of the deficiencies of traditional methods in multivalent rotavirus vaccine potency detection, a cell-based quantitative RT-qPCR assay (C-QPA) was established and validated for specificity, precision, and accuracy. METHODS: In order to further validate the robustness of this method in actual titer detection, the linear range and the practical application under different conditions were tested using monovalent and trivalent rotavirus samples and standards. RESULTS: Results showed that the linear range was 2.0-6.5, 3.9-8.3, and 3.5-8.1 UI (unit of infectivity) for G2, G3, and G4, respectively. Besides, unknown sample with high titer exceeding the linear range can be calculated by dilution. The UIs of serotypes G2, G3, and G4 in monovalent and trivalent rotavirus samples showed a relative deviation ≤4.10%, and the monovalent samples of the same serotype with or without protective agents showed a relative deviation ≤4.28%; the coefficient of variation (CV) of at least 176 tests (548 individual runs) of 3 in vitro-transcribed RNA standards with certain concentrations was not higher than 6.50%; the results of the trivalent samples tested by more than 149 times in 5 years (467 individual runs) showed the CVs lower than 12.66%; 15 samples detected by one laboratory showed a CV lower than 9.83%, while other three samples tested by two independent laboratories showed a CV lower than 6.90%. CONCLUSION: In summary, the C-QPA has good linearity, durability, repeatability, and reproducibility in practical application and has been proved by the authority to be widely used in the production, quality control and release of the recently licensed trivalent vaccine in China.

Evaluation of the diagnostic performance of EpiTuub® Fecal Rotavirus Antigen Rapid Test Kit in Amhara National Regional State, Ethiopia: A multi-center cross-sectional study.

Rotavirus is the leading cause of morbidity and mortality due to acute gastroenteritis among children under five years globally. Early diagnosis of rotavirus infection minimizes its spread and helps to determine the appropriate management of diarrhea. The aim of this study was to evaluate the performance of EpiTuub® Fecal Rotavirus Antigen Rapid Test Kit for the diagnosis of rotavirus infection among diarrheic children under five years in Ethiopian healthcare settings. A total of 537 children with diarrhea were enrolled from three referral hospitals in Amhara National Regional State, Ethiopia. The samples were tested using one-step RT-PCR and EpiTuub® Fecal Rotavirus Antigen Rapid Test Kit (KTR-917, Epitope Diagnostics, San Diego USA) in parallel. Diagnostic performance of the rapid test kit was evaluated using the one-step RT-PCR as a gold standard. The sensitivity, specificity, and predictive values of the rapid test kit were determined. Moreover, the agreement of the rapid test kit with one step RT-PCR was determined by kappa statistics and receiver operators' curve (ROC) analysis was done to assess the overall diagnostic accuracy of the rapid test kit. Fecal Rotavirus Antigen Rapid Test Kit has shown a sensitivity of 75.5% and specificity of 98.2%. The kit was also found to have 89.9% and 95.0% positive and negative predictive values, respectively. The Fecal Rotavirus Antigen Rapid Test Kit has shown a substantial agreement (78.7%, p = 0.0001) with one-step RT-PCR. The overall accuracy of the Fecal Rotavirus Antigen Rapid Test Kit was excellent with the area under the ROC curve of 86.9% (95% CI = 81.6, 92.1%) (p = .0001). Thus, Fecal Rotavirus Antigen Rapid Test is a sensitive, specific, user-friendly, rapid, and equipment-free option to be used at points of care in Ethiopian health care settings where resource is limited precluding the use of one step RT-PCR. Furthermore, the kit could be used in the evaluation and monitoring of rotavirus vaccine effectiveness in the aforementioned settings.

Severe Adult Rotavirus Gastroenteritis: A Rare Case with Multi-Organ Failure and Critical Management.

BACKGROUND Infectious diarrheal illnesses such as rotavirus gastroenteritis are significant contributors to childhood morbidity and mortality, especially in low socio-demographic index regions. Major advances in addressing this issue include sanitation and clean water initiatives, as well as rotavirus immunization. In Australia, a robust vaccination program has significantly reduced childhood rotavirus infections, leading to decreased hospitalizations and mortality. However, cases of adult rotavirus still occur, and although these adult patients usually do not require interventional management, it is possible for them to present critically unwell and require resuscitation. CASE REPORT A previously well 65-year-old man presented to the Emergency Department febrile and hypotensive with severe diarrhea attributed to rotavirus. Clinically, he presented with mixed hypovolemic and septic shock. Despite initial resuscitation, he had multiple severe acute end-organ complications, secondary to poor perfusion. He acquired an acute kidney injury, type-2 myocardial infarction, and ischemic hepatic injury. The mainstay of management was rapid fluid resuscitation, continuous renal replacement therapy, and monitoring in the Intensive Care Unit; however, it was crucial to empirically treat for other causes of shock. CONCLUSIONS To the best of our knowledge, there is a scarcity of reports documenting the management of severe rotavirus gastroenteritis in adults. We recommend advising elderly patients to avoid contact with individuals with diarrheal illnesses, especially rotavirus gastroenteritis. Clinicians should also promote awareness regarding the potential severity of a disease that is typically managed conservatively, and be aware that intervention can be required in severe gastroenteritis.

A Comparison of Pathogen Detection and Risk Factors among Symptomatic Children with Gastroenteritis Compared with Asymptomatic Children in the Post-rotavirus Vaccine Era.

OBJECTIVE: To describe demographics, pathogen distribution/seasonality, and risk factors in children seeking care for acute gastroenteritis (AGE) at a midwestern US emergency department during 5 postrotavirus vaccine years (2011-2016), and further, to compare the same data with matched healthy controls (HC). STUDY DESIGN: AGE and HC participants <11 years old enrolled in the New Vaccine Surveillance Network study between December 2011 to June 2016 were included. AGE was defined as ≥3 diarrhea episodes or ≥1 vomiting episode. Each HC's age was similar to an AGE participant's age. Pathogens were analyzed for seasonality effects. Participant risk factors for AGE illness and pathogen detections were compared between HC and a matched subset of AGE cases. RESULTS: One or more organisms was detected in 1159 of 2503 children (46.3%) with AGE compared with 99 of 537 HC (17.3%). Norovirus was detected most frequently among AGE (n = 568 [22.7%]) and second-most frequently in HC (n = 39 [6.8%]). Rotavirus was the second most frequently detected pathogen among AGE (n = 196 [7.8%]). Children with AGE were significantly more likely to have reported a sick contact compared with HC, both outside the home (15.6% vs 1.4%; P < .001) and inside the home (18.6% vs 2.1%; P < .001). Daycare attendance was higher among children with AGE (41.4%) compared with HC (29.5%; P < .001). The Clostridium difficile detection rate was slightly higher among HC (7.0%) than AGE (5.3%). CONCLUSIONS: Norovirus was the most prevalent pathogen among children with AGE. Norovirus was detected in some HC, suggesting potential asymptomatic shedding among HC. The proportion of AGE participants with a sick contact was approximately 10 times greater than that of HC.

Acute rotavirus infection causes significant activation of the IL-33/IL-13 axis.

AIM: Overactivation of the IL-33/IL-13 axis is the main step in initializing allergic inflammation and promoting allergic diseases. Data on viral pathogens as risk factors for subsequent allergic disease are contradictory. The strongest associations have been made between upper respiratory tract virus infections and asthma. Intestinal viral infections also activate IL-33 and IL-13 as part of the innate antiviral response. The aim of this study was to test whether there are differences in IL-13 and IL-33 concentrations in pediatric patients with acute rotavirus- and norovirus infections and healthy controls. MATERIAL AND METHODS: Forty children with acute rotavirus, 27 with acute norovirus intestinal infections and 17 control children were enrolled in this study. Blood IL-33 and IL-13 detection was performed with enzyme-linked immunosorbent assays (ELISAs). RESULTS: Acute rotavirus infection caused a significant elevation in IL-33 and IL-13 compared to acute norovirus infection (63.85 pg/ml vs. 0, P = 0.0026, and 94.24 pg/ml vs. 0.88 pg/ml, P = 0.0003, respectively) and healthy controls (63.85 pg/ml vs. 9.89 pg/ ml, P = 0.0018, and 94.24 pg/ml vs. 0.14 pg/ml, P < 0.0001, respectively). There was no significant difference in IL-33 and IL-13 concentrations between the acute norovirus group and healthy controls (0 vs. 9.89 pg/ml, P = 0.8276 and 0.88 pg/ml vs. 0.14 pg/ml, P = 0.1652, respectively). CONCLUSION: Acute rotavirus infection causes a significant elevation in IL-33 and IL-13, compared to norovirus and healthy control children.

RT-PCR-based assessment of the SD Bioline Rota/Adeno Antigen-based test in infants with and without diarrhea.

BACKGROUND: Rotavirus A (RVA) infections remain a major cause of severe acute diarrhea affecting children worldwide. To date, rapid diagnostic tests (RDT) are widely used to detect RVA. However, paediatricians question whether the RDT can still detect the virus accurately. Therefore, this study aimed to evaluate the performance of the rapid rotavirus test in comparison to the one-step RT-qPCR method. METHODS: A cross-sectional study was conducted in Lambaréné, Gabon, from April 2018 to November 2019. Stool samples were collected from children under 5 years of age with diarrhoea or a history of diarrhoea within the last 24 h, and from asymptomatic children from the same communities. All stool samples were processed and analysed using the SD BIOLINE Rota/Adeno Ag RDT against a quantitative reverse transcription PCR (RT-qPCR), which is considered the gold standard. RESULTS: For a total of 218 collected stool samples, the overall sensitivity of the RDT was 46.46% (confidence interval (CI) 36.38-56.77), with a specificity of 96.64% (CI 91.62-99.08) compared to one-step RT-qPCR. After confirming the presence or absence of RVA gastroenteritis, the RDT showed suitable results in detecting rotavirus A-associated disease, with a 91% concordance with the RT-qPCR. Furthermore, the performance of this test varied when correlated with seasonality, symptoms, and rotavirus genotype. CONCLUSION: This RDT showed high sensitivity and was suitable for the detection of RVA in patients with RVA gastroenteritis, although some asymptomatic RVA shedding was missed by RT-qPCR. It could be a useful diagnostic tool, especially in low-income countries.

Neurological complications of rotavirus infection in children: A systematic review and meta-analysis.

AIM: To systematically review the clinical features and outcomes of paediatric patients developing neurological complications associated with a rotavirus infection. METHODS: A systematic literature review and meta-analysis was performed, including articles published from 1984 to 2020. Neurological complications were classified into four groups: encephalitis, cerebellitis, encephalo-cerebellitis and benign convulsions with mild gastroenteritis (CwG). RESULTS: Out of 68 reports that fulfilled the research criteria, 99 cases of CwG, 39 cases of encephalitis, 18 cases of encephalo-cerebellitis and five cases of cerebellitis were collected. Ninety-five patients were from Asia. Median age was 22 (IQR 14-29) months, and the children who developed CwG were significantly younger (19, IQR 12-24 months, p < 0.0001) than the others. Status epilepticus was observed in 23% and 5% of the encephalitis and CwG groups respectively. The most frequently described neuroimaging finding were lesions of the splenium of corpus callosum. Four deaths were reported in the encephalitis group, whereas no fatal events were described in the other groups. Among the surviving children, the encephalo-cerebellitis group showed the most severe long-term outcome. All cases of CwG recovered completely. CONCLUSION: Older age at diagnosis and the development of encephalo-cerebellitis are associated with a higher risk of long-term complications.

Diagnostic accuracy of rotavirus antigen tests in children: A systematic review and meta-analysis.

OBJECTIVES: Rotavirus infection is the leading cause of acute gastroenteritis (AGE) in children. Children with rotavirus infection may have symptoms such as diarrhoea or vomiting. Diarrheal diseases caused by rotavirus and other enteric pathogens cannot be differentiated on the basis of clinical symptoms. Therefore, diagnostic testing is essential to confirm a diagnosis of rotavirus infection. The aim of the meta-analysis is to evaluate the diagnostic accuracy of immunochromatographic rotavirus antigen tests in children. METHODS: We searched the PubMed, Embase, Cochrane Library and Google Scholar databases for studies evaluating the diagnostic accuracy of antigen tests for rotavirus in children. We included studies that provided sufficient data to construct a 2 × 2 table on a per patient basis. The overall sensitivity and specificity of the antigen tests were determined using a bivariate random-effects model. RESULTS: In total, 12 studies with 4407 participants were included in the study. The meta-analysis yielded a pooled sensitivity of 89.2% (95% confidence interval [CI]: 77.4%-95.3%) and pooled specificity of 93.2% (95% CI: 83.8%-97.3%). A subgroup analysis of such tests in children aged ≤5 years yielded a pooled sensitivity of 87.1% and pooled specificity of 91.8%. Another subgroup analysis of high-quality studies involving 820 participants yielded a pooled sensitivity of 92.3% and pooled specificity of 95.2%. CONCLUSIONS: Rotavirus antigen tests have high sensitivity for the diagnosis of rotavirus infection in children with AGE. In addition, such tests may be effective for the identification and clinical management of rotavirus infection in children and the prevention of disease progression.

Development and evaluation of polyclonal antibodies based antigen capture ELISA for detection of porcine rotavirus.

Rotaviruses are rising as zoonotic viruses worldwide, causing the lethal dehydrating diarrhea in children, piglets, and other livestock of economic importance. A simple, swift, cost-effective, highly specific, and sensitive antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) was developed for detection of porcine rotavirus-A (PoRVA) by employing rabbit (capture antibody) and murine polyclonal antibodies (detector antibody) produced against VP6 of PoRVA (RVA/Pig-tc/CHN/TM-a/2009/G9P23). Reactivity of the both polyclonal antibodies was confirmed by using an indirect ELISA, western-blot analysis and indirect fluorescence assay against rVP6 protein and PoRVA. The detection limit of AC-ELISA was found 50 ng/ml of PoRVA protein. The relative sensitivity and specificity of this in-house AC-ELISA were evaluated for detection of PoRVA from 295 porcine diarrhea samples, and results were compared with that of RT-PCR and TaqMan RT-qPCR. The relative sensitivity and specificity of AC-ELISA compared with those of TaqMan RT-qPCR were found as 94.4 and 99.2%, respectively, with the strong agreement (κ -0.58) between these two techniques. Furthermore, AC-ELISA could not detect any cross-reactivity with porcine epidemic diarrhea virus, transmissible gastro-enteritis virus, pseudo rabies virus and porcine circovirus-2. This in-house AC-ELISA efficiently detected PoRVA from clinical samples, which suggests that this technique can be used for large-scale surveillance and timely detection of rotavirus infection in the porcine farms.

In this study, we used a Chinese porcine rotavirus-A (PoRVA) strain containing the I5, a dominant VP6-genotype in pigs, for production of VP6 (most conserved) protein based polyclonal antibodies (pAb) in rabbits (as capture Ab) and mouse (as detector Ab) for development of simple, cost effective, highly specific and sensitive AC-ELISA for detection of PoRVA. Furthermore, there is no any previous published report on application of rabbit and mouse pAb against VP6 for developing an AC-ELISA against PoRVA.

Assessment of seasonality of rotavirus PCR detection in swine from Ontario and Quebec between 2016-2020 using submissions to a diagnostic laboratory.

The goal of this study was to determine if seasonality of rotavirus A, B, and C infection is present in Ontario and Quebec swine herds by investigating submissions to a diagnostic laboratory. Samples (N = 1557) within 755 case submissions from Canadian swine herds between 2016 and 2020 were tested for rotaviruses A, B, and C using a real-time polymerase-chain reaction assay and described. Data from Ontario and Quebec were additionally analyzed using boxplots, 6-week rolling averages, time-series decomposition, and negative binomial regression models. Percentage positivity of submissions for rotaviruses A, B, and C were discovered to be highest in nursery/weaner (n = 100, 94.0%, 60.0%, 80.0%) and grower/finisher (n = 13, 84.6%, 46.2%, 61.5%) pigs and lowest in gilt/sow (n = 45, 68.9%, 20.0%, 40.0%) and suckling pigs (n = 102, 67.6%, 10.8%, 38.2%), respectively. The most common combination of rotavirus at the sample level was AC (n = 252, 17%) and ABC (n = 175, 23.2%) at the submission level. Percent positivity for rotavirus A, B, and C across all Canadian provinces included in the study were 69.9%, 32.6%, and 53.1%, respectively. Descriptive analysis suggested little to no evidence of seasonal patterns, although a spike in November was seen in the monthly total submissions and monthly total positive submissions. Statistically, the overall month effect could not be identified as statistically significant (P > 0.05) for any of the evaluated submission counts. Overall, there was no evidence supporting seasonality of rotavirus within Ontario and Quebec swine herds between 2016 and 2020.

Le but de cette étude était de déterminer si la saisonnalité de l'infection à rotavirus A, B et C est présente dans les troupeaux de porcs de l'Ontario et du Québec en examinant les soumissions à un laboratoire de diagnostic. Des échantillons (N = 1557) de 755 cas soumis de troupeaux de porcs canadiens entre 2016 et 2020 ont été testés pour les rotavirus A, B et C à l'aide d'un test de réaction d'amplification en chaîne par polymérase en temps réel et décrits. Les données de l'Ontario et du Québec ont également été analysées à l'aide de diagrammes en boîte, de moyennes mobiles sur 6 semaines, d'une décomposition de séries chronologiques et de modèles de régression binomiale négative. On a découvert que le pourcentage de positivité des soumissions pour les rotavirus A, B et C étaient le plus élevé en pouponnière/sevrage (n = 100, 94,0 %, 60,0 %, 80,0 %) et en croissance/engraissement (n = 13, 84,6 %, 46,2 %, 61,5 %) des porcs et le plus bas chez les cochettes/truies (n = 45, 68,9 %, 20,0 %, 40,0 %) et les porcs à la mamelle (n = 102, 67,6 %, 10,8 %, 38,2 %), respectivement. La combinaison la plus courante de rotavirus au niveau de l'échantillon était AC (n = 252, 17 %) et ABC (n = 175, 23,2 %) au niveau de la soumission. Les pourcentages de positivité pour les rotavirus A, B et C dans toutes les provinces canadiennes incluses dans l'étude étaient de 69,9 %, 32,6 % et 53,1 %, respectivement. L'analyse descriptive a suggéré peu ou pas de preuves de tendances saisonnières, bien qu'un pic en novembre ait été observé dans les soumissions totales mensuelles et les soumissions positives totales mensuelles. Statistiquement, l'effet mensuel global n'a pu être identifié comme statistiquement significatif (P > 0,05) pour aucun des nombres de soumissions évalués. Dans l'ensemble, il n'y avait aucune preuve à l'appui de la saisonnalité du rotavirus dans les troupeaux de porcs de l'Ontario et du Québec entre 2016 et 2020.(Traduit par Docteur Serge Messier).

Human rotavirus VP4 and VP7 genetic diversity and detection of GII norovirus in Ibadan as Nigeria introduces rotavirus vaccine.

OBJECTIVE: This cross-sectional study investigated the circulating strains of rotavirus and screened for noravirus in Ibadan, Nigeria as the country introduces the rotavirus vaccine into its national immunization program. METHODS: Sixty-five stool samples were collected from children younger than 5 years with clinically diagnosed diarrhea and screened for the presence of rotavirus and norovirus using RT-PCR. Rotavirus-positive samples were further analyzed to determine the G and P genotypes using semi-nested multiplex PCR. RESULTS: The rates of rotavirus and norovirus positivity were 30.8% and 10.8%, respectively, whereas the rate of rotavirus and norovirus mixed infection was 4.6%. G1 was the predominant VP7 genotype, followed by G2, G9, and G1G2G9, whereas the predominant VP4 genotype was P[4], followed by P[6], P[8], and P[9]. The mixed P types P[4]P[8] and P[4]P[6] were also detected. G1P[4] was the most common VP4 and VP7 combination, followed by G2P[4], G1[P6], G1P[8], G2P[6], G2P[9], G9P[6], G2G9P[4], G2P[4]P[6], G1P[4]P[8], G2G9P[8], G1G2G9P[8], and G1[non-typable] P[non-typable], which were detected in at least 5% of the samples. Four samples had a combination of non-typable G and P types. CONCLUSIONS: It is essential to monitor the circulation of virus strains prior to and during the implementation of the immunization program.

Norovirus-associated White Matter Injury in a Term Newborn With Seizures.

Neonatal seizures with white matter injury have been associated with rotavirus, enterovirus and parechovirus. Neurological symptoms caused by norovirus have been occasionally reported in older children. We describe a case of a neonate with seizures and white matter lesions, with detection of human norovirus in stool samples from the patient and her mother.

Development of a rapid homogeneous immunoassay for detection of rotavirus in stool samples.

Rotavirus is the main pathogen causing acute viral gastroenteritis. Accurate and rapid diagnosis of rotavirus infection is important to determine appropriate treatment, prevention of unnecessary antibiotics use and control of infection spread. In this study, we established a rapid, accurate, and sensitive amplified luminescent proximity homogeneous assay linked immunosorbent assay (AlphaLISA) for detecting rotavirus and evaluated its efficacy in human stool samples. Our results demonstrated that the sensitivity of AlphaLISA (5-8) significantly exceeded that of the immunochromatographic assay (ICA, 5-4) for rotavirus antigen detection. The intra-assay and inter-assay coefficients of variation were 2.99-3.85% and 5.27-6.51%, respectively. Furthermore, AlphaLISA was specific for rotavirus and did not cross-react with other common diarrhea viruses. AlphaLISA and real-time reverse transcription polymerase chain reaction (RT-qPCR, which is considered a gold standard for detecting diarrhea viruses) tests showed consistent results on 235 stool samples, with an overall consistency rate of 97.87% and a kappa value of 0.894 (P < 0.001). The overall consistency rate of ICA compared with RT-qPCR was 95.74%. AlphaLISA showed better consistency with RT-qPCR than the routinely used ICA for rotavirus detection in stool samples. The AlphaLISA method can be used in clinical practice for the rapid, accurate, and sensitive detection of rotavirus infection.

Rotavirus and Norovirus Infections in Children Under 5 Years Old with Acute Gastroenteritis in Southwestern China, 2018-2020.

OBJECTIVE: Rotaviruses and noroviruses are important causes of acute gastroenteritis in children. While previous studies in China have mainly focused on rotavirus, we investigated the incidence of norovirus in addition to rotavirus in Southwestern China. METHODS: From January 2018 to December 2020, cases of rotavirus or norovirus infections among children under five ages with acute gastroenteritis were evaluated retrospectively. RESULTS: The detection rate of rotavirus was 24.5% (27,237/111,070) and norovirus was 26.1% (4649/17,797). Among 17,113 cases submitted for dual testing of both rotavirus and norovirus, mixed rotavirus/norovirus infections were detected in 5.0% (859/17,113) of cases. While there was no difference in norovirus incidence in outpatient compared to hospitalized cases, rotavirus was detected two times more in outpatients compared to hospitalized cases (26.6% vs.13.6%; P < 0.001). Both rotavirus and norovirus infections peaked in children aged 12-18 months seeking medical care with acute gastroenteritis (35.6% rotavirus cases; 8439/23,728 and 32.5% norovirus cases; 1660/5107). Rotavirus infections were frequent between December and March of each year while norovirus was detected earlier from October to December. Our results showed significant correlation between virus detection and environmental factors such as average monthly temperature but not relative humidity. In addition, we observed a reduction in the detection rates of rotavirus and norovirus at the beginning of the SARS-CoV-2 pandemic in 2020. CONCLUSION: Our results indicate that rotavirus and norovirus are still important viral agents in pediatric acute gastroenteritis in Southwestern China.