Rapid differential detection of classical and highly pathogenic North American Porcine Reproductive and Respiratory Syndrome virus in China by a duplex real-time RT-PCR

2009 ◽  
Vol 161 (2) ◽  
pp. 192-198 ◽  
Author(s):  
Nan-Hua Chen ◽  
Xi-Zhao Chen ◽  
Dong-Mei Hu ◽  
Xiu-Ling Yu ◽  
Li-Lin Wang ◽  
...  
Keyword(s):  
Real Time ◽  
North American ◽  
Respiratory Syndrome Virus ◽  
Differential Detection ◽  
Rt Pcr ◽  
Highly Pathogenic

scholarly journals Simultaneous Detection of North American and European Porcine Reproductive and Respiratory Syndrome Virus Using Real-Time Quantitative Reverse Transcriptase–PCR

2005 ◽  
Vol 17 (2) ◽  
pp. 165-170 ◽  
Author(s):  
Steven B. Kleiboeker ◽  
Susan K. Schommer ◽  
Sang-Myeong Lee ◽  
Sandy Watkins ◽  
Wayne Chittick ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
North American ◽  
Simultaneous Detection ◽  
Viral Rna ◽  
Respiratory Syndrome Virus ◽  
Analytical Sensitivity ◽  
Rt Pcr ◽  
Field Isolates ◽  
Pcr Assays

Porcine reproductive and respiratory syndrome (PRRS) is 1 of the most economically important diseases of swine. Detection of the etiologic agent, PRRS virus (PRRSV), represents a diagnostic challenge due to the heterogeneity of field isolates as well as the propensity for swine to develop persistent infection in which virus is difficult to detect. Recently European (EU) lineage PRRSV isolates, which are genetically divergent from North American (NA) isolates, have been introduced into NA swine further complicating efforts to diagnose this disease. In this study, real-time ( TaqMan) reverse transcriptase (RT)–PCR assays were developed for multiplex detection, differentiation, and quantification of NA and EU PRRSV field isolates. Oligonucleotide primers and dual-labeled probes were selected from conserved regions of open-reading frame 7 and the 3'-untranslated region. The real-time RT-PCR assays described for the NA or EU genotype of PRRSV detected viral RNA from 83/83 strains (74 NA; 9 EU) previously isolated by cell culture between 1992 and 2003. The analytical sensitivity of both assays was consistently found to be less than a single TCID50, which corresponded to 5–10 RNA molecules, and was not significantly reduced when the reactions were performed in a multiplex format. When performing multiplex reactions, sensitive detection was possible even when 1 viral RNA concentration was up to 5,000-fold higher than the second. The diagnostic sensitivity and specificity of the multiplex reaction was found to be at a minimum equivalent to that of both nested RT-PCR and virus isolation.

scholarly journals Detection and Typing of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus by Multiplex Real-Time RT-PCR

PLoS ONE ◽  
2012 ◽  
Vol 7 (6) ◽  
pp. e38251 ◽  
Author(s):  
Kerstin Wernike ◽  
Bernd Hoffmann ◽  
Malte Dauber ◽  
Elke Lange ◽  
Horst Schirrmeier ◽  
...  
Keyword(s):  
Real Time ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
Highly Pathogenic

Validation and comparison of different end point and real time RT-PCR assays for detection and genotyping of porcine reproductive and respiratory syndrome virus

2014 ◽  
Vol 201 ◽  
pp. 79-85 ◽  
Author(s):  
Michele Drigo ◽  
Giovanni Franzo ◽  
Ilaria Belfanti ◽  
Marco Martini ◽  
Alessandra Mondin ◽  
...  
Keyword(s):  
Real Time ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
End Point ◽  
Pcr Assays

scholarly journals Sequencing and analysis of complete genome of the attenuated Hanvet1.VN strain used for vaccine production against porcine reproductive and respiratory syndrome

2018 ◽  
Vol 16 (1) ◽  
pp. 51-57
Author(s):  
Nguyễn Thị Nga ◽  
Hà Thị Thu ◽  
Nguyễn Thị Hoa ◽  
Vũ Thị Hiền ◽  
Trần Thị Thu Hiền ◽  
...  
Keyword(s):  
North American ◽  
Sanger Sequencing ◽  
Respiratory Syndrome Virus ◽  
Whole Genome ◽  
Type Ii ◽  
Vaccine Production ◽  
Rt Pcr ◽  
North American Strain ◽  
American Strain ◽  
Vietnamese Strain

The porcine reproductive and respiratory syndrome virus (PRRSV) attenuated strain Hanvet1.VN has been developed by the Pharmaceutical and Veterinary Material J.S.C (HANVET) by passaging HY-2010 strain on MARC-145 cells for 80 passages and used for PRRS vaccine production. In this study, we sequenced and analyzed the whole genome of the attenuated Hanvet1.VN strain. The total RNA was extracted from the Hanvet1.VN strain, RT-PCR was used for amplification of 15 separate segments of the whole genome. The amplified segments were cloned into the pCR2.1 vector and sequenced by Sanger sequencing. The sequences were analyzed with BioEdit and DNA Star Software. The results showed that, GP5 of the Hanvet1.VN attenuated strain had 100% identity in amino acid (aa) sequences with one of the pathogenic Vietnamese strain isolated in Quang Nam Province and had 98% identity with that of the Chinese 07NM strain. However, the identity of aa sequence of the Hanvet1.VN GP5 was much lower in the comparison with GP5 of VR2332, and it was only 87%. The MP and NP proteins were highly conserved compared with pathogenic strains circulating in Vietnam (07QN) and China (07NM) (99-100%, respectively). The other eight proteins of the Hanvet1.VN strain showed changes from 1.2% in NP1a to 3.9% in GP2 compared with the 07QN strain. However, the aa identity of all Hanvet1.VN proteins were very low when compared with proteins of PRRSV type II strain (North American strain, VR2332), ranged from 86.25% to 97.7%. Our results showed that the Hanvet1.VN attenuated vaccine strain had protective immunogenicity similar to that strain circulating in Vietnam closely related to a strain from China but different from the type II North American strain VR2332. Hence, for importing PRRSV vaccine, especially from American or Europe Countries, antigenic compatibility of the PRRSV vaccine and strains circulating in Vietnam should be concerned in PRRSV vaccine production.

scholarly journals High frequency RNA recombination in porcine reproductive and respiratory syndrome virus occurs preferentially between parental sequences with high similarity

2001 ◽  
Vol 82 (11) ◽  
pp. 2615-2620 ◽  
Author(s):  
Joke J. F. A. van Vugt ◽  
Torben Storgaard ◽  
Martin B. Oleksiewicz ◽  
Anette Bøtner
Keyword(s):  
North American ◽  
High Frequency ◽  
Respiratory Syndrome Virus ◽  
European American ◽  
Rna Recombination ◽  
High Similarity ◽  
Rt Pcr ◽  
European Isolate ◽  
North American Type ◽  
Theoretical Risk

Two types of porcine reproductive and respiratory syndrome virus (PRRSV) exist, a North American type and a European type. The co-existence of both types in some countries, such as Denmark, Slovakia and Canada, creates a risk of inter-type recombination. To evaluate this risk, cell cultures were co-infected with either a North American and a European type of PRRSV or two diverse types of European isolate. Subsequently, an approximately 600 bp region of the PRRSV genome was tested for recombination by quantitative real-time RT–PCR. Between 0·1 and 2·5% RNA recombination was found between the European isolates, but no recombination was detected between the European and North American types. Calculation of the maximum theoretical risk of European–American recombination, based on the sensitivity of the RT–PCR system, revealed that RNA recombination between the European and North American types of PRRSV is at least 10000 times less likely to occur than RNA recombination between diverse European isolates.

scholarly journals Development of a real-time RT-PCR method for the detection of newly emerged highly pathogenic H7N9 influenza viruses

2017 ◽  
Vol 16 (9) ◽  
pp. 2055-2061 ◽  
Author(s):  
Xiu-rong WANG ◽  
Lin-lin GU ◽  
Jian-zhong SHI ◽  
Hai-feng XU ◽  
Ying ZHANG ◽  
...  
Keyword(s):  
Real Time ◽  
Influenza Viruses ◽  
Rt Pcr ◽  
Highly Pathogenic ◽  
Pcr Method ◽  
H7n9 Influenza
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