Multiplex PCR and multiplex RT-PCR for inclusive detection of major swine DNA and RNA viruses in pigs with multiple infections

Ogawa, Hirohito; Taira, Osamu; Hirai, Takuya; Takeuchi, Hiromi; Nagao, Aki; Ishikawa, Yoshiki; Tuchiya, Kotaro; Nunoya, Tetsuo; Ueda, Seinosuke

doi:10.1016/j.jviromet.2009.05.010

Cited by 80 publications

(55 citation statements)

References 24 publications

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“…The multi-site amplification is carried out in the same condition and reaction system, there are amplifying competition and mutual influence among different primers. Besides, change of temperature , G+C content and specificity of primers (Ogawa et al 2009, Durzyńska et al 2011) may also affect the results. For better results, the primers of this experiment were designed carefully.…”

Section: Discussionmentioning

confidence: 99%

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Hu¹,

Lin²,

Yang³

et al. 2015

Polish Journal of Veterinary Sciences

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In this assay, we developed and evaluated a multiplex PCR (mPCR) for its ability in detecting multiple infections of swine simultaneously. Four pairs of primers were used to detect five viruses. Specific primers were designed for classical swine fever virus (CSFV), African swine fever virus (ASFV) and pseudorabies (PRV). A pair of primers was designed prudently for two different types of porcine reproductive and respiratory syndrome virus that respectively were porcine reproductive and respiratory syndrome virus (PRRSV), highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV). The detection limits of the mPCR were 1. A total of 49 clinical specimens were tested by the mPCR, and the result showed that co-infection by two or three viruses was 51%. In conclusion, the PCR is a useful tool for clinical diagnosis of not only single infections but also mixed infections in swines.

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Section: Discussionmentioning

confidence: 99%

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Hu¹,

Lin²,

Yang³

et al. 2015

Polish Journal of Veterinary Sciences

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“…In the polymerase chain reaction (PCR) to amplify the PCV2 genome primers used were: forward 5'-CAGCAACATGCCCAGCAAGAAGAAT-3' and reverse 5'-TCGATCACACAGTCTCAGTAG-3' (Ogawa et al, 2009), which flank a fragment of 703 bp from the PCV2 ORF V1. The components of the reaction mix were: PCR buffer 1x to make a final volume of 25 µl, MgCl2 1.5 mM, nucleotide mix, d'NTPs 0.2 mM, each primer 20 pmol, Taq Gold DNA polymerase 1.25 U, and sample DNA 2 µl (0.1-50 ng).…”

Section: Amplification and Detection Of Pcv2 Dnamentioning

confidence: 99%

Study of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) frequencies and coinfection in mexican farrow to finish pig farms

Vargas¹,

Escatell²,

Lara³

et al. 2018

J. Vet. Med. Anim. Health

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The porcine circovirus type 2 (PCV2) is commonly present along with the porcine reproductive and respiratory syndrome virus (PRRSV). These are the most important pathogens for pig industry worldwide. In this study, we determined frequencies for both viruses, as well as coinfection, in farrow to finish farms from México. For this, pigs from 28 farms in different states were sampled for tonsils, lungs and lymphatic nodes and assayed for PCV2 and PRRSV by polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR). Herds detected positive were: 16 PCV2 (57%), 10 PRRSV (36%); and 5 had both viruses (18%). Samples detected positives were: 49% PCV2, and 39% PRRSV. Coinfection frequency observed in this work contrasted those observed in other countries. We think that these data will contribute to a better understanding of both diseases in order to take better measures for preventing and controlling them.

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“…Other diagnostic techniques, such as conventional PCR or real -time PCR are widely used [12]. Different studies showed that PCR is very sensitive method for PPV detection [13].…”

Section: Introductionmentioning

confidence: 99%

Molecular Detection of PCV2 And PPV in Pigs in Republic of Srpska, Bosnia and Herzegovina

et al. 2016

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The presence of porcine circovirus 2 and porcine parvovirus was examined in forty clinical samples of spleen, lymph nodes and lungs originating from non-vaccinated swine by polymerase chain reaction. All animals were reared in extensive livestock farming systems in different geographical districts of Republic of Srpska, Bosnia and Herzegovina. Porcine circovirus 2 DNA was detected in four lymph node and two spleen samples (15%), while porcine parvovirus DNA was identifi ed in fi ve lymph node samples (12.5%). The presence of both viruses was detected in three lymph node samples (7.5%). Partial nucleotide sequence of ORF1 gene of 2 porcine circovirus 2 and VP2 gene of 2 porcine parvovirus isolates was determined. The nucleotide sequences of two PCV2 isolates from RS-BIH included in phylogenetic typing are similar and cluster together with the strain Mantova isolated from domestic pigs in Italy, strains DE006-14 and DE222-13 isolated from pigs in Germany as well as with the strain Jvnan isolated from pigs in China. Also, analyzed PCV2 isolates were partially similar to the strain NIV-C SRB isolated from pigs in Serbia. The nucleotide sequences of two PPV isolates that were included in phylogenetic typing showed a high level of similarity with the strain Challenge isolated from pigs in UK, strain Kresse isolated from pigs in USA and strains 77 and LZ isolated from pigs in China.

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Multiplex PCR and multiplex RT-PCR for inclusive detection of major swine DNA and RNA viruses in pigs with multiple infections

Cited by 80 publications

References 24 publications

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

A multiplex PCR for simultaneous detection of classical swine fever virus, African swine fever virus, highly pathogenic porcine reproductive and respiratory syndrome virus, porcine reproductive and respiratory syndrome virus and pseudorabies in swines

Study of porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV) frequencies and coinfection in mexican farrow to finish pig farms

Molecular Detection of PCV2 And PPV in Pigs in Republic of Srpska, Bosnia and Herzegovina

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