Development and Application of a Multiplex PCR Method for Rapid Differential Detection of Subgroup A, B, and J Avian Leukosis Viruses

Gao, Qi; Yun, Bingling; Wang, Q.; Jiang, Lili; Zhu, Huabin; Gao, Yulong; Qin, Liting; Wang, Y.; Qi, Xiaole; Gao, Honglei; Wang, Xifeng

doi:10.1128/jcm.02200-13

Cited by 52 publications

(41 citation statements)

References 38 publications

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“…The mPCR method is based on a single PCR method. Therefore, although this method has the advantages of simple operation, time savings, and low cost, there are inherent weaknesses of sensitivity due to the amount of competitive inhibition between the amplicons, which renders the sensitivity of the mPCR lower than that of single PCR [1,4,16]. Therefore, our expectation of the mPCR effect was that when the viral titer of each virus in the sample exceeded the sensitivity of the mPCR method, all viruses would be detected.…”

Section: Discussionmentioning

confidence: 99%

“…The multiplex PCR (mPCR) method is based on a single PCR that can simultaneously detect a variety of pathogens and simplify the operation steps, thereby saving time and expense [3]. Recently, mPCR has been widely used in the field of pathogen detection because it has advantages of speed, specificity and efficiency [1,4,16]. In this study, an mPCR method was established and applied for the detection of CPV-2, CAV and CCoV.…”

Section: Introductionmentioning

confidence: 99%

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A multiplex PCR method for the simultaneous detection of three viruses associated with canine viral enteric infections

Deng

Zhang

et al. 2018

Arch Virol

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The aim of this study was to establish a multiplex PCR (mPCR) method that can simultaneously detect canine parvovirus (CPV-2), canine coronavirus (CCoV) and canine adenovirus (CAV), thereby eliminating the need to detect these pathogens individually. Based on conserved regions in the genomes of these three viruses, the VP2 gene of CPV-2, the endoribonuclease nsp15 gene of CCoV, and the 52K gene of CAV were selected for primer design. The specificity of the mPCR results showed no amplification of canine distemper virus (CDV), canine parainfluenza virus (CPIV), or pseudorabies virus (PRV), indicating that the method had good specificity. A sensitivity test showed that the detection limit of the mPCR method was 1 × 10 viral copies. A total of 63 rectal swabs from dogs with diarrheal symptoms were evaluated using mPCR and routine PCR. The ratio of positive samples to total samples for CPV-2, CCoV, and CAV was 55.6% (35/63) for mPCR and 55.6% (35/63) for routine PCR. Thirty-five positive samples were detected by both methods, for a coincidence ratio of 100%. This mPCR method can simultaneously detect CCoV (CCoV-II), CAV (CAV-1, CAV-2) and CPV-2 (CPV-2a, CPV-2b, CPV-2c), which are associated with viral enteritis, thereby providing an efficient, inexpensive, specific, and accurate new tool for clinical diagnosis and laboratory epidemiological investigations.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A multiplex PCR method for the simultaneous detection of three viruses associated with canine viral enteric infections

Deng

Zhang

et al. 2018

Arch Virol

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“…1 We also detected other viruses using PCRs specific for avian reovirus (ARV), avian leukosis virus (ALV), Marek's disease virus (MDV), reticuloendotheliosis virus (REV), CAV and avian adenovirus (ADV). 10, 11, 12 All positive PCR products were sequenced for verification.…”

mentioning

confidence: 99%

Avian gyrovirus 2 in poultry, China, 2015–2016

Yao

Tuo

Gao

et al. 2016

Emerging Microbes & Infections

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“…PCR that used to detect other pathogens including MDV (F: 5 0 -T GCGATGAAAGTGCTATGGAGG-3 0 ; R: 5 0 -GAGAATCCCTATGAGAAAGCGC-3 0 ), AGV2 (F: 5 0 -CGTGTCCGCCAGCAGAAAC-3 0 ; R: 5 0 -GGTAGAAGCCAAAGCGTCCAC-3 0 ) [9], ALV (F: 5 0 -GGATGAGGTGACTAAGAAAG-3 0 ; R: 5 0 -CGAACCAAAGGTAACAC ACG-3 0 ) [21], REV (F: 5 0 -GCCTTAGCCGCCATTGTA-3 0 ; R: 5 0 -CCAGCCAACACC ACGAACA-3 0 ) [22] and ARV (F: 5 0 -ATGGCGGGTCTCAATCCATC-3 0 ; R: 5 0 -TTA GGTGTCGATGCCGGTAC-3 0 ) [23].…”

Section: Amplification Of the Cav Genomementioning

confidence: 99%

Molecular epidemiology of chicken anaemia virus in sick chickens in China from 2014 to 2015

et al. 2019

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Chicken anaemia virus (CAV), a member of the genus Gyrovirus, is the etiological agent of chicken infectious anaemia. CAV infects bone marrow-derived cells, resulting in severe anaemia and immunosuppression in young chickens and a compromised immune response in older birds. We investigated the molecular epidemiology of CAV in sick chickens in China from 2014 to 2015 and showed that the CAV-positive rate was 13.30%, in which mixed infection (55.56%) was the main type of infection. We isolated and identified 15 new CAV strains using different methods including indirect immunofluorescence assay and Western Blotting. We used overlapping polymerase chain reaction to map the whole genome of the strains. Phylogenetic analyses of the obtained sequences and related sequences available in GenBank generated four distinct groups (A–D). We built phylogenetic trees using predicted viral protein (VP) sequences. Unlike CAV VP2s and VP3s that were well conserved, the diversity of VP1s indicated that the new strains were virulent. Our epidemiological study provided new insights into the prevalence of CAV in clinical settings in recent years in China.

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Development and Application of a Multiplex PCR Method for Rapid Differential Detection of Subgroup A, B, and J Avian Leukosis Viruses

Cited by 52 publications

References 38 publications

A multiplex PCR method for the simultaneous detection of three viruses associated with canine viral enteric infections

A multiplex PCR method for the simultaneous detection of three viruses associated with canine viral enteric infections

Avian gyrovirus 2 in poultry, China, 2015–2016

Molecular epidemiology of chicken anaemia virus in sick chickens in China from 2014 to 2015

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