scholarly journals Diarrhea caused by rotavirus A, B, and C in suckling piglets from southern Brazil: molecular detection and histologic and immunohistochemical characterization

2018 ◽  
Vol 30 (3) ◽  
pp. 370-376 ◽  
Author(s):  
Paula R. Almeida ◽  
Elis Lorenzetti ◽  
Raquel S. Cruz ◽  
Tatiane T. Watanabe ◽  
Priscila Zlotowski ◽  
...  
Keyword(s):  
Virus Detection ◽  
Enteric Virus ◽  
Transmissible Gastroenteritis Virus ◽  
Southern Brazil ◽  
Rt Pcr ◽  
Viral Pathogen ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Suckling Piglets ◽  
Transmissible Gastroenteritis

Rotavirus (RV) is an important viral pathogen causing diarrhea in piglets and other mammals worldwide. We describe 34 cases from 4 diarrheal outbreaks caused by RV in unvaccinated farrowing units in southern Brazil from 2011 to 2013. We performed autopsy, histologic examinations, bacterial culture, RV immunohistochemistry (IHC), and enteric virus detection through molecular assays for rotavirus A, B, and C, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, sapovirus, norovirus, and kobuvirus. Histologically, villus atrophy (29 of 34) and epithelial vacuolation (27 of 34) occurred in all 4 outbreaks. Cell debris in the lamina propria occurred in 20 cases, mostly from outbreaks A (8 of 11), C (4 of 6), and D (7 of 11). IHC was positive for RV in 21 of 34 samples. RT-PCR was positive for RV in 20 of 30 samples; RV-C was the most frequently detected RV ( n = 17). Kobuvirus was detected in 11 samples, and, in 3 of them, there was single detection of this enteric virus.

THE OUTBREAK OF PORCINE EPIDEMIC DIARRHEA IN TAIWAN

2014 ◽  
Vol 40 (03) ◽  
pp. 115-121 ◽  
Author(s):  
Ming-Chung Deng ◽  
Chia-Yi Chang ◽  
Tien-Shine Huang ◽  
Shu-Ting Kuo ◽  
Hsiang-Jung Tsai ◽  
...  
Keyword(s):  
Animal Health ◽  
Vero Cells ◽  
Transmissible Gastroenteritis Virus ◽  
Rt Pcr ◽  
Sequence Comparisons ◽  
Diarrhea Virus ◽  
Severe Diarrhea ◽  
Porcine Epidemic Diarrhea ◽  
Transmissible Gastroenteritis ◽  
Group 2

Between January 20 and April 30 of 2014, a total of 103 diarrhea cases from 47 herds in 13 counties were submitted to the Animal Health Research Institute. In 20 of the 25 herds with detail history, severe diarrhea and vomiting occurred in pigs of all ages, with mortality approaching 100% in suckling pigs. The differential etiologies, including transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), and porcine group A rotavirus (GARV), were tested by reverse transcription polymerase chain reaction (RT-PCR). The RT-PCR of PEDV was positive in 79 cases of 34 herds. Attempts to isolate PEDV in Vero cells revealed that only 7 specimens from 7 herds showed the cytopathic effects (CPEs) of fusion and syncytia. These CPEs were indeed caused by PEDV, as confirmed by RT-PCR, sequencing, and electron microscopy. Sequence comparisons of diarrhea samples and isolated PEDV were assayed by MEGA 5.2 software. The newly isolated PEDV/Taiwan/2014 strains were clustered in group 2 as novel PEDV, together with strains PEDV/USA/2013, PEDV/China/2011–2013, PEDV/Thailand/2007–2008, and PEDV/Korea/2008–2009, whereas the classical CV777 strain was placed in a separate group 1. These results indicated that a novel PEDV was the cause of the recent new outbreak of diarrhea in Taiwan.

scholarly journals Use of dual priming oligonucleotide system-based multiplex RT-PCR assay to detect five diarrhea viruses in pig herds in South China

AMB Express ◽  
2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Guangbin Si ◽  
Jiawei Niu ◽  
Xia Zhou ◽  
Yongsheng Xie ◽  
Zhifei Chen ◽  
...  
Keyword(s):  
Detection Efficiency ◽  
Limit Of Detection ◽  
Transmissible Gastroenteritis Virus ◽  
Rt Pcr ◽  
Simple Method ◽  
Detection Rates ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Dual Priming Oligonucleotide ◽  
Pcr Method

AbstractIn this study, a specific and simple method based on the dual priming oligonucleotide (DPO) system was developed to simultaneously detect transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), porcine rotavirus A (PRV-A), porcine delta coronavirus (PDCoV), and swine acute diarrhea syndrome coronavirus (SADS-CoV), associated with the major enteric RNA viruses in pigs. The DPO system-based multiplex RT-PCR method simplified the primer design and did not require optimization of the annealing temperature. Specificity analysis revealed that the method could specifically detect TGEV, PEDV, PRV-A, PDCoV, and SADS-CoV without any cross-amplification of other circulating swine viruses. The limit of detection of the method was as low as 103–104 copies/μL plasmid of each virus. The method also had good repeatability, and obvious results were seen in three repeat experiments with an interval of 45 days. This optimized multiplex RT-PCR method was used to evaluate 181 clinical swine samples that were collected from four provinces of China between September 2016 and August 2018. The results showed that the positive detection rates of PEDV, PDCoV, SADS-CoV, PRV-A, and TGEV were 30.94% (56/181), 17.67% (32/181), 11.6% (21/181), 9.39% (17/181), and 0.55% (1/181), respectively. Mixed infection of two or more viruses was also common. The DPO system-based multiplex RT-PCR could be a useful tool for detecting enteric virus infections. This method has the advantages of easy operation, low cost, high detection efficiency, and short running time for early diagnosis in clinical cases.

scholarly journals Use of dual priming oligonucleotide system-based multiplex RT-PCR assay to detect five diarrhea viruses in pig herds in South China

2021 ◽  
Author(s):  
Guangbin Si ◽  
Jiawei Niu ◽  
Xia Zhou ◽  
Yongsheng Xie ◽  
Zhifei Chen ◽  
...  
Keyword(s):  
Detection Efficiency ◽  
Limit Of Detection ◽  
Transmissible Gastroenteritis Virus ◽  
Rt Pcr ◽  
Simple Method ◽  
Detection Rates ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Dual Priming Oligonucleotide ◽  
Pcr Method

Abstract In this study, a specific and simple method based on the dual priming oligonucleotide (DPO) system was developed to simultaneously detect transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), porcine rotavirus A (PRV-A), porcine delta coronavirus (PDCoV), and swine acute diarrhea syndrome coronavirus (SADS-CoV), associated with the major enteric RNA viruses in pigs. The multiplex RT-PCR method based on the DPO system simplified the primer design and did not require optimization of the annealing temperature. Specificity analysis revealed that the method could specifically detect TGEV, PEDV, PRV-A, PDCoV, and SADS-CoV without any cross-amplification of other circulating swine viruses. The limit of detection of the method was as low as 103–104 copies/μL plasmid of each virus. The method also had good repeatability, and obvious results were seen in three repeat experiments with an interval of 45 days. This optimized multiplex RT-PCR method was used to evaluate 181 clinical swine samples that were collected from four provinces of China between September 2016 and August 2018. The results showed that the positive detection rates of PEDV, PDCoV, SADS-CoV, PRV-A, and TGEV were 30.94% (56/181), 17.67% (32/181), 11.6% (21/181), 9.39% (17/181), and 0.55% (1/181), respectively. Mixed infection of two or more viruses was also common. The DPO system-based multiplex RT-PCR could be a useful tool for detecting enteric virus infections. This method has the advantages of easy operation, low cost, high detection efficiency, and short running time for early diagnosis in clinical cases.

scholarly journals Data standardization implementation and applications within and among diagnostic laboratories: integrating and monitoring enteric coronaviruses

2021 ◽  
pp. 104063872110021
Author(s):  
Giovani Trevisan ◽  
Leticia C. M. Linhares ◽  
Kent J. Schwartz ◽  
Eric R. Burrough ◽  
Edison de S. Magalhães ◽  
...  
Keyword(s):  
Seasonal Pattern ◽  
Laboratory Data ◽  
Information Management System ◽  
Transmissible Gastroenteritis Virus ◽  
Test Results ◽  
Diarrhea Virus ◽  
Oral Fluids ◽  
Transmissible Gastroenteritis ◽  
Technology Processes ◽  
Laboratory Information Management

Every day, thousands of samples from diverse populations of animals are submitted to veterinary diagnostic laboratories (VDLs) for testing. Each VDL has its own laboratory information management system (LIMS), with processes and procedures to capture submission information, perform laboratory tests, define the boundaries of test results (i.e., positive or negative), and report results, in addition to internal business and accounting applications. Enormous quantities of data are accumulated and stored within VDL LIMSs. There is a need for platforms that allow VDLs to exchange and share portions of laboratory data using standardized, reliable, and sustainable information technology processes. Here we report concepts and applications for standardization and aggregation of data from swine submissions to multiple VDLs to detect and monitor porcine enteric coronaviruses by RT-PCR. Oral fluids, feces, and fecal swabs were the specimens submitted most frequently for enteric coronavirus testing. Statistical algorithms were used successfully to scan and monitor the overall and state-specific percentage of positive submissions. Major findings revealed a consistently recurrent seasonal pattern, with the highest percentage of positive submissions detected during December–February for porcine epidemic diarrhea virus, porcine deltacoronavirus, and transmissible gastroenteritis virus (TGEV). After 2014, very few submissions tested positive for TGEV. Monitoring VDL data proactively has the potential to signal and alert stakeholders early of significant changes from expected detection. We demonstrate the importance of, and applications for, data organized and aggregated by using LOINC and SNOMED CTs, as well as the use of customized messaging to allow inter-VDL exchange of information.

scholarly journals Multiplex Detection of Rotavirus A, B, C, Norovirus GI and GII, Adenovirus, Astrovirus and Sapovirus in a Single PCR System in Stool Samples From Acute Diarrhea Children

2020 ◽  
Author(s):  
Wei Li ◽  
Weiwei Li ◽  
Ran Tao ◽  
Wenqing Xiang ◽  
Mingming Zhou ◽  
...  
Keyword(s):  
High Throughput ◽  
Acute Diarrhea ◽  
Cross Reaction ◽  
Melting Curve Analysis ◽  
Clinical Test ◽  
Rt Pcr ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Intestinal Pathogens ◽  
Stool Samples

Abstract Background: Early and accurate identification of infection viruses among children can benefit the personalized medical treatment and management, and reduce the future occurrence of serious symptoms. Thus, it is critical to develop a high-throughput multiplex real-time RT-PCR method to improve the accuracy and efficiency in routine clinical lab tests. Here, we developed a RT-PCR combined with melting curve analysis (RRCMC) method for simultaneous detection of rotavirus A, B, C, norovirus GI and GII, adenovirus, astrovirus and sapovirus. Results: Stool samples were collected from 160 children with acute diarrhea and tested by RRCMC assay. A total of 71 patients were tested positive with norovirus, adenovirus or rotavirus. The RRCMC assay has high specificity. There is no internal cross-reaction through the 8 diarrhea viruses and no cross-reaction of other commonly intestinal pathogens and human genome. The detection limit was ranging from 1×102 to 1×105 copies/ml for each diarrhea virus. Conclusions: In conclusion, the RRCMC method is a suitable rapid clinical test for infection viruses, with the advantages of high-throughput, low cost, high sensitivity, and specificity.

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