Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection

Camargos, Marcelo Fernandes; Stancek, D; Lessa, Leandro Moreira; Reis, Jenner Karlisson Pimenta dos; Rocha, Maurilio Andrade; Leite, Rômulo Cerqueira

doi:10.1590/s1413-95962003000500005

Cited by 1 publication

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“…On the other hand, PCR detected BLV proviral DNA in blood from 9 animals that tested negative in all the serologic tests. Those results show, again, a disparity between direct and indirect tests (Camargos et al, 2003).…”

Section: Dementioning

confidence: 87%

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Testes de diagnóstico para o vírus da leucemia bovina

Camargos¹,

Júnior²,

Cruz³

et al. 2005

RBCV

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Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis (EBL). After infection with BLV there is no detectable viremia but there is a strong and persistent humoral immune response to structural proteins, essentially the gp51 envelope glycoprotein and the major core protein p24. Polymerase chain reaction (PCR) with primers used to amplify part of env gene, agar gel immunodiffusion (AGID) which employs a crude antigen preparation derived from concentrated cell culture fluid, two commercial immuno enzymatic assays (ELISAs) and one ELISA with recombinant gp51 antigen were used to detect proviral DNA and BLV antibodies in DNA and serum from bovines of Goias (GO), Mato Grosso do Sul (MS), Minas Gerais (MG) and Paraná (PR) States of Brazil. The evaluated tests presented good results. The ELISA tests were more sensitive than AGID, and highly specific. PCR sensitivity should be improved, nevertheless it is a good complementary tool for serologic diagnosis. Resumo: O Vírus da Leucemia Bovina (BLV) é o agente etiológico da Leucose Enzoótica Bovina (LEB). Após a infecção não se observa viremia, mas desenvolve-se uma resposta imune humoral forte e duradoura às proteínas estruturais, principalmente a glicoproteína do envelope gp51, do envelope, e à proteína do cerne p24. A reação em cadeia pela polimerase (PCR), com iniciadores direcionados à amplificação de fragmentos do gene env do BLV, a imunodifusão em gel de agar (IDGA), que emprega antígeno bruto derivado de sobrenadante de cultivo celular concentrado, dois ensaios imunoenzimáticos (ELISA) comerciais e um ELISA que emprega a proteína gp51 recombinante foram utilizados para detectar, respectivamente, o DNA proviral e anticorpos para o BLV em amostras de DNA e soro de bovinos dos estados de Goiás, Mato Grosso do Sul, Minas Gerais e Paraná. Os testes de ELISA foram mais sensíveis que a IDGA e altamente específicos. A PCR revelou-se útil como ferramenta de diagnóstico complementar aos testes sorológicos.

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Section: Dementioning

confidence: 87%

“…The PCR was done as described by Camargos et al (2003), using primers of BLV env gene to amplify fragments with 521 pairs of bases (bp): BLV1 5'-GGGCCATGGTCACATATGATTG-3' (5128-5149) e BLV2 5'-CGTTGCCTTGAGAAACATTGAAC-3' (5627-5649). The numbers correspond to those of BLV sequences published by Sagata et al (1985).…”

Section: Dementioning

confidence: 99%