Development of a real-time RT-qPCR assay for the detection of porcine respirovirus 1

Li, Yanhua; Sthal, Chase; Bai, Jianfa; Liu, Xuming; Anderson, Gary A.; Fang, Ying

doi:10.1016/j.jviromet.2020.114040

Cited by 11 publications

(9 citation statements)

References 7 publications

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“…Although the RT‐qPCR assay did reveal positive detection in samples not previously tested by SMg, it could not confirm PPIV‐1 detection in all SMg‐positive samples. A recently published paper designed a PPIV‐1 RT‐qPCR assay with primers based on the NCBI GenBank sequences, but contrary to our design, targeted the hemagglutinin–neuraminidase gene (Li et al., 2021 ). This RT‐qPCR approach should be able to detect the strains described in our study (in silico prediction).…”

Section: Resultsmentioning

confidence: 99%

“…Nevertheless, PPIV‐1 was only detected within 1/32 BS, compared to 2/4 NS and 8/32 OF samples. Additionally, in a study performed by Li and colleagues, RT‐qPCR analysis of 49 BS samples of PPIV‐1 infected pigs yielded no positive BS results (Li et al., 2021 ). As a result, it would suggest a limited suitability for detecting PPIV‐1 in BS, compared to NS and OF samples.…”

Section: Resultsmentioning

confidence: 99%

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First detection of porcine respirovirus 1 in Germany and the Netherlands

Schuele

Lizarazo-Forero

Cassidy

et al. 2021

Transbound Emerg Dis

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Porcine respirovirus 1, also referred to as porcine parainfluenza virus 1 (PPIV-1), was first detected in deceased pigs from Hong Kong in 2013. It has since then been found in the USA, Chile and most recently in Hungary. Information on the pathogenicity and global spread is sparse. However, it has been speculated to play a role in the porcine respiratory disease complex. To investigate the porcine virome, we screened 53 pig samples from 26 farms within the Dutch-German border region using shotgun metagenomics sequencing (SMg). After detecting PPIV-1 in five farms through SMg, a real-time reverse transcriptase PCR (RT-qPCR) assay was designed, which not only confirmed the presence of the virus in 1 of the 5 farms but found an additional 6 positive farms. Phylogenetic analysis found the closest match to be the first detected PPIV-1 strain in Hong Kong. The Dutch-German region represents a significant area of pig farming within Europe and could provide important information on the characterization and circulation of porcine viruses, such as PPIV-1. With its recent detection in Hungary, these findings suggest widespread circulation of PPIV-1 in Central Europe, highlighting the need for further research on persistence, pathogenicity and transmission in Europe.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

First detection of porcine respirovirus 1 in Germany and the Netherlands

Schuele

Lizarazo-Forero

Cassidy

et al. 2021

Transbound Emerg Dis

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“…RNA was extracted using QIAamp cador Pathogen Mini Kit (Qiagen, Hilden, Germany), according to the manufacturer's instruction. Extracted RNA (4 µL) was used for RT real-time PCR with SensiFAST Probe No-ROX One Step Kit Kit (Bioline, London, UK), with primers and a probe targeting a highly conserved region in H-N gene [12]. The amplification was carried out using 6000 Rotor Gene (Qiagen, Hilden, Germany) under the following thermal conditions: 45 • C/10 min, 95 • C/10 min, followed by 40 cycles of 95 • C/15 s, 56 • C/20 s, and 72 • C/20 s. Samples with a cycle threshold (Ct) ≤ 37.0 were considered positive.…”

Section: Detection Of Prv1 Iav and Prrsv In Clinical Samplesmentioning

confidence: 99%

Detection of Porcine Respirovirus 1 (PRV1) in Poland: Incidence of Co-Infections with Influenza A Virus (IAV) and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in Herds with a Respiratory Disease

Woźniak

Cybulski²,

Dénes

et al. 2022

Viruses

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Porcine respirovirus 1 (PRV1) is also known as porcine parainfluenza virus 1 (PPIV1). The prevalence and the role of PRV1 infections for pig health is largely unknown. In order to assess the PRV1 prevalence in Poland, nasal swabs and oral fluids collected from pigs from 30 farms were examined with RT real-time PCR. Additionally, IAV and PRRSV infection statuses of PRV1-positive samples were examined. The results showed that the virus is highly prevalent (76.7% farms positive) and different patterns of PRV1 circulation in herds with mild–moderate respiratory disease were observed. Co-infections with IAV and PRRSV were infrequent and detected in 8 (23.5%) and 4 (11.8%) out of 34 PRV1-positive nasal swab pools from diseased pens, respectively. In one pen PRV1, IAV, and PRRSV were detected at the same time. Interestingly, PRV1 mean Ct value in samples with co-infections was significantly lower (29.8 ± 3.1) than in samples with a single PRV1 infection (32.5 ± 3.6) (p < 0.05), which suggested higher virus replication in these populations. On the other hand, the virus detection in pig populations exhibiting respiratory clinical signs, negative for PRRSV and IAV, suggests that PRV1 should be involved in differential diagnosis of respiratory problems.

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“…The samples originated from 12 farms, 7 of which belonged to a single company ( Table 1 ) [ 8 ]. The samples that tested positive in real-time RT-PCR with Ct < 30 [ 14 ] were used in RT-PCR in order to generate amplicons encompassing the whole F gene of PPIV-1, which were then sequenced with the Sanger method ( Supplementary Table S1 ). The obtained DNA sequences were assembled with Geneious 10.2.6 (Biomatters Ltd., Auckland, New Zealand).…”

Section: Methodsmentioning

confidence: 99%

European and American Strains of Porcine Parainfluenza Virus 1 (PPIV-1) Belong to Two Distinct Genetic Lineages

Stadejek

Cybulski²,

Gauger

et al. 2022

Pathogens

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Porcine parainfluenza virus 1 (PPIV-1) is a recently emerged respirovirus closely related to human parainfluenza virus 1 (HPIV-1) and Sendai virus (SenV). PPIV-1 has been detected in Asia, the Americas and Europe, but knowledge on its epidemiology and genetic diversity is very limited. In the present study, the complete nucleotide sequences of the fusion (F)-protein gene obtained from samples from 12 Polish and 11 US herds were analysed and compared to previously available genetic data from the Americas, Asia and Europe. The existence of two distinct clades was observed, grouping European sequences and one Hong Kong sequence (clade 1), or one American sequence and three Asian sequences (clade 2). The mean genetic distances measured with the p-distance were 0.04 (S.E., 0.000) within both clades, and 0.095 (S.E., 0.006) between the clades. Moreover, two distinct clusters of highly similar sequences were identified, which corresponded to the geographically distant nurseries and finishing units, from three pig flows within one Polish pig-production company. The obtained data indicate that the two PPIV-1 lineages may have evolved independently in Europe and America. More studies, particularly involving Asian viruses, are necessary to understand the virus’ emergence and epidemiology and the role of carriers in the spread of PPIV-1.

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Development of a real-time RT-qPCR assay for the detection of porcine respirovirus 1

Cited by 11 publications

References 7 publications

First detection of porcine respirovirus 1 in Germany and the Netherlands

First detection of porcine respirovirus 1 in Germany and the Netherlands

Detection of Porcine Respirovirus 1 (PRV1) in Poland: Incidence of Co-Infections with Influenza A Virus (IAV) and Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) in Herds with a Respiratory Disease

European and American Strains of Porcine Parainfluenza Virus 1 (PPIV-1) Belong to Two Distinct Genetic Lineages

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