Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China
Abstract Background Diarrheal diseases caused by viral agents have led to a great morbidity, mortality, and economic loss in global pig industry. Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and group A porcine rotavirus (RVA)...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-03-01
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| Series: | Porcine Health Management |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s40813-024-00364-y |
| _version_ | 1797272777884958720 |
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| author | Wenbo Song Yixue Feng Jiali Zhang Danni Kong Jie Fan Mengfei Zhao Lin Hua Jinmei Xiang Xibiao Tang Shaobo Xiao Zhong Peng Bin Wu |
| author_facet | Wenbo Song Yixue Feng Jiali Zhang Danni Kong Jie Fan Mengfei Zhao Lin Hua Jinmei Xiang Xibiao Tang Shaobo Xiao Zhong Peng Bin Wu |
| author_sort | Wenbo Song |
| collection | DOAJ |
| description | Abstract Background Diarrheal diseases caused by viral agents have led to a great morbidity, mortality, and economic loss in global pig industry. Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and group A porcine rotavirus (RVA) are main causative agents of swine viral diarrhea with similar clinical signs on Chinese farms and their co-infection is also common. However, it is still lack of a convenient method to detect these four agents. Methods A TaqMan multiplex qPCR method was developed to detect PEDV, TGEV, PDCoV, and RVA, simultaneously. This method was then applied to investigate 7,342 swine fecal samples or rectal swabs, as well as 1,246 swine intestinal samples collected from 2075 farms in China in 2022. Results Minimum detection limits of this method were 3 copies/µL for PEDV, 4 copies/µL for TGEV, 8 copies/µL for RVA, and 8 copies/µL for PDCoV, suggesting a good sensitivity. No signals were observed by using this method detecting other viral agents commonly prevalent in pigs, which is suggestive of a good specificity. Application of this method on investigating clinical samples demonstrated a relatively high positive rate for PEDV (22.21%, 1907/8588) and RVA (44.00%, 3779/8588). In addition, co-infection between PEDV and RVA was observed on 360 investigated farms, accounting for 17.35% (360/2075) of the farms where co-infection events were screened. Conclusions A TaqMan multiplex qPCR method targeting PEDV, TGEV, PDCoV, and RVA was developed in this study. This method demonstrated a good specificity and sensitivity on investigating these four common viruses responsible for viral diarrhea on Chinese pig farms, which represents a convenient method for the monitoring and differential diagnosis of swine viral diarrhea. |
| first_indexed | 2024-03-07T14:34:14Z |
| format | Article |
| id | doaj.art-aa3c4dd572b146b599e7e8156eb90791 |
| institution | Directory Open Access Journal |
| issn | 2055-5660 |
| language | English |
| last_indexed | 2024-03-07T14:34:14Z |
| publishDate | 2024-03-01 |
| publisher | BMC |
| record_format | Article |
| series | Porcine Health Management |
| spelling | doaj.art-aa3c4dd572b146b599e7e8156eb907912024-03-05T20:44:12ZengBMCPorcine Health Management2055-56602024-03-011011910.1186/s40813-024-00364-yDevelopment of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in ChinaWenbo Song0Yixue Feng1Jiali Zhang2Danni Kong3Jie Fan4Mengfei Zhao5Lin Hua6Jinmei Xiang7Xibiao Tang8Shaobo Xiao9Zhong Peng10Bin Wu11National Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityAnimal Disease Diagnosis Center of Wuhan Keqian Biology Co., LtdAnimal Disease Diagnosis Center of Wuhan Keqian Biology Co., LtdAnimal Disease Diagnosis Center of Wuhan Keqian Biology Co., LtdNational Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityNational Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityNational Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityHubei Vocational College of Bio-TechnologyAnimal Disease Diagnosis Center of Wuhan Keqian Biology Co., LtdNational Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityNational Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityNational Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityAbstract Background Diarrheal diseases caused by viral agents have led to a great morbidity, mortality, and economic loss in global pig industry. Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine deltacoronavirus (PDCoV), and group A porcine rotavirus (RVA) are main causative agents of swine viral diarrhea with similar clinical signs on Chinese farms and their co-infection is also common. However, it is still lack of a convenient method to detect these four agents. Methods A TaqMan multiplex qPCR method was developed to detect PEDV, TGEV, PDCoV, and RVA, simultaneously. This method was then applied to investigate 7,342 swine fecal samples or rectal swabs, as well as 1,246 swine intestinal samples collected from 2075 farms in China in 2022. Results Minimum detection limits of this method were 3 copies/µL for PEDV, 4 copies/µL for TGEV, 8 copies/µL for RVA, and 8 copies/µL for PDCoV, suggesting a good sensitivity. No signals were observed by using this method detecting other viral agents commonly prevalent in pigs, which is suggestive of a good specificity. Application of this method on investigating clinical samples demonstrated a relatively high positive rate for PEDV (22.21%, 1907/8588) and RVA (44.00%, 3779/8588). In addition, co-infection between PEDV and RVA was observed on 360 investigated farms, accounting for 17.35% (360/2075) of the farms where co-infection events were screened. Conclusions A TaqMan multiplex qPCR method targeting PEDV, TGEV, PDCoV, and RVA was developed in this study. This method demonstrated a good specificity and sensitivity on investigating these four common viruses responsible for viral diarrhea on Chinese pig farms, which represents a convenient method for the monitoring and differential diagnosis of swine viral diarrhea.https://doi.org/10.1186/s40813-024-00364-yTaqMan multiplex qPCR methodPEDVTGEVPDCoVRVASwine viral diarrhea |
| spellingShingle | Wenbo Song Yixue Feng Jiali Zhang Danni Kong Jie Fan Mengfei Zhao Lin Hua Jinmei Xiang Xibiao Tang Shaobo Xiao Zhong Peng Bin Wu Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China Porcine Health Management TaqMan multiplex qPCR method PEDV TGEV PDCoV RVA Swine viral diarrhea |
| title | Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China |
| title_full | Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China |
| title_fullStr | Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China |
| title_full_unstemmed | Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China |
| title_short | Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China |
| title_sort | development of a multiplex reverse transcription quantitative pcr qpcr method for detecting common causative agents of swine viral diarrhea in china |
| topic | TaqMan multiplex qPCR method PEDV TGEV PDCoV RVA Swine viral diarrhea |
| url | https://doi.org/10.1186/s40813-024-00364-y |
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