Determination of the Detection Limit of the Polymerase Chain Reaction for Chicken Infectious Anemia Virus

Soiné, Christiane; Watson, SJ; Rybicki, Edward P.; Lucio, Benjamin; Nordgren, Robert; Parrish, Colin R.; Schat, Karel A.

doi:10.2307/1591674

Cited by 33 publications

(16 citation statements)

References 24 publications

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“…10,20,22,24 Although the use of nested PCR showed improved detection sensitivity by 10-100-fold, 10,20 it increases the cost and time required for vaccine evaluation with no extra positive impact on the results obtained.…”

Section: Discussionmentioning

confidence: 99%

An Optimized Polymerase Chain Reaction Assay to Identify Avian Virus Vaccine Contamination with Chicken Anemia Virus

et al. 2011

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Abstract. The use of embryonating chicken eggs in preparation of avian virus vaccines is the principle cause for contamination with Chicken anemia virus (CAV). Identification of CAV in contaminated vaccines relies on the expensive, tedious, and time-consuming practice of virus isolation in lymphoblastoid cell lines. The experience of the last 2 decades indicates that polymerase chain reaction is extending to replace most of the classic methods for detection of infectious agents. In the present report, a simple, rapid, and accurate polymerase chain reaction method for detection of CAV in poultry vaccines is described. Oligonucleotide primers homologous to highly conserved sequences of the VP1 gene were used to amplify a fragment of 676 bp. The developed assay was specific for detecting CAV from different sources, with no cross reactivity with many avian viruses. No inter-and intra-assay variations were observed. The analytical sensitivity of the test was high enough to detect 5 TCID 50 (50% tissue culture infective dose) of the virus per reaction; however, different factors related to the vaccine matrix showed considerable effects on the detection limit. In conclusion, this method may represent a suitable alternative to virus isolation for identification of CAV contamination of poultry virus vaccines.

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Section: Discussionmentioning

confidence: 99%

An Optimized Polymerase Chain Reaction Assay to Identify Avian Virus Vaccine Contamination with Chicken Anemia Virus

et al. 2011

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Section: Discussionmentioning

confidence: 99%

“…Another PCR (Noteborn et al, 1992a,b) and a nested-PCR (Soiné et al, 1993) were also reported only for the detection of CAV in MSB-1 cell cultures infected with CAV rather than directly in chicken tissues from field cases suspected of CAV, and had already indicated as well that the PCR assay is highly sensitive for the diagnosis of CAV. The sensitivity of the nested-PCR from Soiné et al (1993) was evaluated in regard to the amplification of cloned CAV DNA and could detect as little as one calculated virus genome equivalent, suggesting that the nested-PCR is an even more sensitive and thus more suitable test for CAV diagnosis than PCR. However, due to its sensitivity, the authors indicate that the nested-PCR is much more prone to cross contamination and therefore requires extreme caution in handling field samples and has to be carried preferably in separate facilities and equipment (Soiné et al, 1993), a matter that has to be even better observed when a nested-PCR is considered for routine use in diagnostic laboratories.…”

Section: Discussionmentioning

confidence: 99%

A VP3/VP1 gene polymerase chain reaction assay for detection of chicken anemia virus in broiler samples

Nogueira

Brentano

Ferreira

2005

Arq. Bras. Med. Vet. Zootec.

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A PCR assay was designed for amplification of the highly conserved VP3 gene and a 5' region of the VP1 gene, for the diagnosis of CAV in organ samples of broiler flocks suspected of chicken infectious anemia. A comparison of the VP3/VP1 PCR with in vivo virus isolation revealed 100% agreement of the results, with 13 positive and 3 negative samples in both assays, indicating that the VP3/VP1 PCR is a specific diagnostic method. Tissues from additional 24 broiler chicken flocks, with CAV-like lesions and clinical history were then tested only by the VP3/VP1 PCR and a reference PCR with published primers for the VP1 gene. Nineteen samples resulted positive and one negative in both PCR, while another 4 samples were positive only in the VP3/VP1 PCR. These results indicate that the VP3/VP1 PCR is a sensitive, specific diagnostic test, suitable as an alternative to the expensive and time consuming in vivo virus isolation method, specially considering the difficult diagnosis of CAV strains not readily adaptable to MSB-1 cell culture.

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“…Two oligonucleotide primers flanking fragment A were as follows. Forward primer (CAV5): 5  -ATC GAA TTC CGA GTG GTT ACT ATT CC -3  (nt 2317-23) and the reverse primer (CAV6): 5  -GAA GGA TCC CTC ATT CTT AGT GGC -3  (nt 1515-1492) (Soiné et al, 1993).…”

Section: Amplification Of Dna Fragments By Polymerase Chain Reaction mentioning

confidence: 99%

Restriction endonuclease analysis of the genomes of different isolates of chicken anemia virus amplified by polymerase chain reaction

et al. 2018

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Four DNA fragments (fragments A, B, C and D) covering the whole genome of chicken anemia virus (CAV) were amplified enzymatically by polymerase chain reaction (PCR) using four pairs of oligonucleotide primers. The DNA fragments were amplified from each of nine CAV isolates including eight Malaysian isolates and one European isolate (Cux-1). For all nine CAV isolates, fragment A (1518 bp) was digested with one restriction enzyme, Eco130I (StyI); fragment B (926 bp) with three enzymes, Eco130I (StyI), HpaII and MboI separately; fragment C (675 bp) with also three enzymes, BsuRI (HaeIII), HinfI, and HpaII separately; and the fragment D (552 bp) with one enzyme, EcoRI. Enzyme digested products of different fragments were separated by agarose gel or polyacrylamide gel electrophoresis. Each of the eightenzymatic reactions differentiated at least two isolates except the HpaII digestion of fragment C where no isolate was distinguished. The overall restriction endonuclease (RE) analysis separated four isolates (BL-1, BL-2, BL-4 and BL-5) in one group and the rest five isolates (SMSC-1, SMSC-2, 3-1, BL-3 and Cux-1) were differentiated from each other and also from the group of four isolates, based on the number of restriction site differences and the fragments generated by different enzymatic digestions. The study revealed that RE analysis could be used to identify and differentiate CAV isolates based on the number of restriction site differences. The study showed that more isolates, even the isolates from the same poultry farm, could be differentiated with proper genomic diversity after RE analysis of more genome fragments compared to that of single genome fragment.

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Determination of the Detection Limit of the Polymerase Chain Reaction for Chicken Infectious Anemia Virus

Cited by 33 publications

References 24 publications

An Optimized Polymerase Chain Reaction Assay to Identify Avian Virus Vaccine Contamination with Chicken Anemia Virus

An Optimized Polymerase Chain Reaction Assay to Identify Avian Virus Vaccine Contamination with Chicken Anemia Virus

A VP3/VP1 gene polymerase chain reaction assay for detection of chicken anemia virus in broiler samples

Restriction endonuclease analysis of the genomes of different isolates of chicken anemia virus amplified by polymerase chain reaction

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