Direct Detection of Bovine Leukemia Virus Infection: Practical Applicability of a Double Polymerase Chain Reaction*

Ballagi-Pordány, A.; Klintevall, K.; Mérza, M.; Klingeborn, B.; Belák, Sándór

doi:10.1111/j.1439-0450.1992.tb01140.x

Cited by 37 publications

(27 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The majority of the PCR assays are based on a single assay. However, it has been reported that less than eight genome copies of the provirus could be detected in the background of 2 million negative lymphocytes using nested PCR (13). Therefore, this study used primers designed for 9 segments of different sizes, based on the envelope gene (GenBank AY078387) of the BLV, and a primer set consisting of both the BLV-2 and BLV-8 was used for the nested PCR.…”

Section: Discussionmentioning

confidence: 99%

Investigation of the Bovine Leukemia Virus Proviral DNA in Human Leukemias and Lung cancers in Korea

et al. 2005

View full text Add to dashboard Cite

The bovine leukemia virus (BLV) is the causative agent of enzootic bovine leucosis. This study investigated the presence of the BLV in leukemia (179 acute lymphoblastic leukemia, 292 acute myeloid leukemia and 46 chronic myelogenous leukemia cases) and 162 lung cancer patients (139 adenocarcinoma, 23 squamous cell carcinoma) to determine if the BLV is a causative organism of leukemia and lung cancer in Koreans. A BLV infection was confirmed in human cells by PCR using a BLV-8 primer combination. All 517 cases of human leukemia and 162 lung cancer were negative for a PCR of the BLV proviral DNA. In conclusion, although meat has been imported from BLV endemic areas, the BLV infection does not appear to be the cause of human leukemia or lung cancer in Koreans. These results can be used as a control for further studies on the BLV in Koreans.

show abstract

Section: Discussionmentioning

confidence: 99%

Investigation of the Bovine Leukemia Virus Proviral DNA in Human Leukemias and Lung cancers in Korea

et al. 2005

View full text Add to dashboard Cite

show abstract

“…The BLV env gene encodes a precursor protein that is processed into two subunits, gp51, the outer membrane subunit, and gp30, the transmembrane subunit, both of which are essential for viral infectivity [20, 28]. A variety of diagnostic tests have been developed for BLV, including PCR-based assays, agar gel immunodiffusions (AGIDs), virus neutralization assays, and enzyme-linked immunosorbent assays (ELISAs) [1,9,11,12,16,25]. The most widely used diagnostic methods for the serological detection of BLV-specific antibodies are AGID and ELISA, which are antigen-based assays that use either wild type BLV gp51 isolated from fetal lamb kidney (FLK) cells, or a recombinant BLV antigen that is expressed in insect cells [6,16,19,23].…”

Section: Introductionmentioning

confidence: 99%

Agar gel immunodiffusion analysis using baculovirus-expressed recombinant bovine leukemia virus envelope glycoprotein (gp51/gp30T-)

et al. 2009

View full text Add to dashboard Cite

Bovine leukemia virus (BLV) envelope glycoprotein (gp51/gp30T-), consisting of BLV gp51 and BLV gp30 that lacked its C-terminal transmembrane domain, was expressed in insect cells under the control of the baculovirus polyhedron promoter. Recombinant BLV gp51/gp30T- secreted from insect cells was determined by immunofluorescence, enzyme-linked immunosorbent and western blot assays using a BLV-specific monoclonal antibody and BLV-positive bovine antibodies. An agar gel immunodiffusion (AGID) test using gp51/gp30T- as the antigen for the detection of BLV antibodies in serum was developed and compared to traditional AGID, which uses wild type BLV antigen derived from fetal lamb kidney cells. AGID with the recombinant BLV gp51/gp30T- was relatively more sensitive than traditional AGID. When the two methods were tested with bovine sera from the field, the recombinant BLV gp51/gp30T- and traditional antigen had a relative sensitivity of 69.8% and 67.4%, respectively, and a relative specificity of 93.3% and 92.3%. These results indicated that the recombinant BLV gp51/gp30T- is an effective alternative antigen for the diagnosis of BLV infection in cattle.

show abstract

“…However, the application of PCR for identification of viral DNA has provided a sensitive test for detecting BLV in tissues such as blood. 1,2,10 This report describes the application of PCR to the detection of BLV DNA in peripheral blood leukocytes and semen of bulls. Such analysis should facilitate the international movement of germ plasm derived from seropositive animals.…”

mentioning

confidence: 99%

Absence of Bovine Leukosis Virus in Semen of Seropositive Bulls

2002

View full text Add to dashboard Cite

Abstract.A polymerase chain reaction (PCR)-based detection system was established to identify the presence of bovine leukosis virus (BLV) DNA in bovine semen. Seventy-nine bulls were included in the study. Serum, peripheral blood leukocytes, and semen were collected from each of the 79 bulls. The BLV-specific antibody was detected in serum by agar gel immunodiffusion and viral DNA in blood and semen by PCR. Serologically, 29 of the 79 bulls were BLV positive. Twenty-seven of the 29 seropositive bulls and 1 of the seronegative bulls had BLV DNA in peripheral blood leukocytes. All 79 bulls tested PCR negative for the presence of BLV in semen. This data is strong evidence that properly collected semen from BLV seropositive bulls will not contribute to dissemination of this viral infection.Bovine leukosis virus (BLV) is an exogenous oncogenic retrovirus associated with enzootic bovine leukosis. The infection can result in 3 different clinical states, with many animals remaining asymptomatic in an aleukemic state (AL). However, 30-70% develop a persistent lymphocytosis (PL) characterized by a polyclonal expansion of B-lymphocytes and a much smaller percent, 0.1-10%, eventually develop lymphoid tumors. 3,6,11,12 A previous survey published in 1985 suggested up to 30% of dairy cows in the United States were infected with BLV. 12 Due to the relatively short life of production cattle in the United States, the greatest economic impact of BLV infection would appear to be that associated with interference in the international movement of cattle and their germ plasm rather than clinical disease. Thus, application of molecular biology techniques, such as polymerase chain reaction (PCR), to identify the presence of BLV in semen should facilitate certification of germ plasm as being free of contaminating virus.Previous studies by numerous investigators suggested that semen from BLV-infected bulls was noninfectious for recipient cows. 4,8,9 Sheep inoculated with entire ejaculates from BLV-infected bulls remained seronegative. Cows inseminated with ejaculates from seropositive bulls have also not seroconverted nor have their progeny. In addition, a 5-year study demonstrated that use of semen derived from both BLV seropositive and seronegative donor bulls in a BLV-free closed dairy did not result in any seroconversion. Donor bulls were from multiple artificial insemination (AI) centers, with 48% being seropositive. 9 Despite the above-mentioned studies that argued strongly against the possibility of recipient cows becoming infected by using semen from BLV-positive bulls, international movement of germ plasm from BLV-infected animals is still problematic. The most common method of testing for BLV in animals is the detection of antibodies to gp51 by agar gel immunodiffusion (AGID). However, the application of PCR for identification of viral DNA has provided a sensitive test for detecting BLV in tissues such as blood. 1,2,10 This report describes the application of PCR to the detection of BLV DNA in peripheral blood leukocytes and se...

show abstract

Direct Detection of Bovine Leukemia Virus Infection: Practical Applicability of a Double Polymerase Chain Reaction*

Cited by 37 publications

References 23 publications

Investigation of the Bovine Leukemia Virus Proviral DNA in Human Leukemias and Lung cancers in Korea

Investigation of the Bovine Leukemia Virus Proviral DNA in Human Leukemias and Lung cancers in Korea

Agar gel immunodiffusion analysis using baculovirus-expressed recombinant bovine leukemia virus envelope glycoprotein (gp51/gp30T-)

Absence of Bovine Leukosis Virus in Semen of Seropositive Bulls

Contact Info

Product

Resources

About