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Reverse Transcription Cross-Priming Amplification-Nucleic Acid Test Strip for Rapid Detection of Porcine Epidemic Diarrhea Virus.

Wang FX, Yuan DY, Jin YN, Hu L, Sun ZY, He Q, Zhao SH, Zhan SB, Wen YJ - Sci Rep (2016)

Bottom Line: The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens.It was also successfully applied to detecting PEDV in clinical specimens.The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, No. 4899 Juye Avenue, Jingyue Economic and Technological Development Zone, Changchun, Jilin, 130112, People's Republic of China.

ABSTRACT
Porcine epidemic diarrhea virus (PEDV) is a highly transmissible coronavirus that causes a severe enteric disease particularly in neonatal piglets. In this study, a rapid method for detecting PEDV was developed based on cross-priming amplification and nucleic acid test strip(CPA-NATS). Five primers specific for the N gene sequence of PEDV were used for the cross-priming amplification. Detection of amplification products based on labeled probe primers was conducted with strip binding antibody of labeled markers. The CPA method was evaluated and compared with a PCR method. The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens. Results showed that the method was highly specific for the detection of PEDV, and had the same sensitivity as PCR, with detection limit of 10(-6) diluted plasmid containing the target gene of PEDV. It was also successfully applied to detecting PEDV in clinical specimens. The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine.

No MeSH data available.


Related in: MedlinePlus

Trial of RT CPA-NATS detecting clinical specimens: (a) result of PEDV RT CPA-NATS and (b) result of PEDV RT-PCR.
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f4: Trial of RT CPA-NATS detecting clinical specimens: (a) result of PEDV RT CPA-NATS and (b) result of PEDV RT-PCR.

Mentions: Additionally, the system was validated by technicians with limited laboratory trainings on a pig farm. Six fecal swab specimens from sick piglets showing disease symptoms Suspected infected by PEDV were collected and tested for PEDV. Two of the six specimens (2# and 3#) were tested positive (Fig. 4a), which was in agreement with the results of RT-PCR (Fig. 4b).


Reverse Transcription Cross-Priming Amplification-Nucleic Acid Test Strip for Rapid Detection of Porcine Epidemic Diarrhea Virus.

Wang FX, Yuan DY, Jin YN, Hu L, Sun ZY, He Q, Zhao SH, Zhan SB, Wen YJ - Sci Rep (2016)

Trial of RT CPA-NATS detecting clinical specimens: (a) result of PEDV RT CPA-NATS and (b) result of PEDV RT-PCR.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4835727&req=5

f4: Trial of RT CPA-NATS detecting clinical specimens: (a) result of PEDV RT CPA-NATS and (b) result of PEDV RT-PCR.
Mentions: Additionally, the system was validated by technicians with limited laboratory trainings on a pig farm. Six fecal swab specimens from sick piglets showing disease symptoms Suspected infected by PEDV were collected and tested for PEDV. Two of the six specimens (2# and 3#) were tested positive (Fig. 4a), which was in agreement with the results of RT-PCR (Fig. 4b).

Bottom Line: The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens.It was also successfully applied to detecting PEDV in clinical specimens.The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Special Economic Animal Molecular Biology, Institute of Special Economic Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, No. 4899 Juye Avenue, Jingyue Economic and Technological Development Zone, Changchun, Jilin, 130112, People's Republic of China.

ABSTRACT
Porcine epidemic diarrhea virus (PEDV) is a highly transmissible coronavirus that causes a severe enteric disease particularly in neonatal piglets. In this study, a rapid method for detecting PEDV was developed based on cross-priming amplification and nucleic acid test strip(CPA-NATS). Five primers specific for the N gene sequence of PEDV were used for the cross-priming amplification. Detection of amplification products based on labeled probe primers was conducted with strip binding antibody of labeled markers. The CPA method was evaluated and compared with a PCR method. The reverse transcription CPA system was further optimized for detecting PEDV RNA in clinical specimens. Results showed that the method was highly specific for the detection of PEDV, and had the same sensitivity as PCR, with detection limit of 10(-6) diluted plasmid containing the target gene of PEDV. It was also successfully applied to detecting PEDV in clinical specimens. The reverse transcription CPA-NATS detection system established in this study offers a specific, sensitive, rapid, and simple detection tool for screening PEDV, which can contribute to strategies in the effective control of PEDV in swine.

No MeSH data available.


Related in: MedlinePlus