A novel real time PCR assay for bovine leukemia virus detection using mixed probes and degenerate primers targeting novel BLV strains

Borjigin, Liushiqi; Yoneyama, Shuji; Saito, Susumu; Polat, Meripet; Inokuma, Michihito; Shinozaki, Yasuo; Tanaka, Naoko; Yamanaka, Risa; Yasui, Anna; Mimura, Munehito; Murakami, Hironobu; Takeshima, Shin‐nosuke; Aida, Yoko

doi:10.1016/j.jviromet.2021.114264

Cited by 3 publications

(3 citation statements)

References 46 publications

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“…To investigate the effect of BoLA-DRB3 polymorphism on milk from BLV-infected dams, we first collected peripheral blood from 672 Holstein-Friesian cattle on eight dairy farms throughout Japan to determine the presence of BLV infection (Table 1). DNA was extracted from blood samples to measure the PVL of known and novel BLV variants in infected cattle using the BLV-CoCoMo-qPCR-2 assay (RIKEN Genesis, Kanagawa, Japan) [34][35][36][37]. In total, 300 of 672 cattle (44.6%) were positive for BLV PVL, and the BLV-positivity rate ranged from 14.7 to 71.7% among the farms (Table 1).…”

Section: Blv Infection Rate and Frequencies Of Susceptible Neutral An...mentioning

confidence: 99%

BoLA-DRB3 Polymorphism Controls Proviral Load and Infectivity of Bovine Leukemia Virus (BLV) in Milk

et al. 2022

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Bovine leukemia virus (BLV), which causes enzootic bovine leukosis, is transmitted to calves through the milk of BLV-infected dams. Bovine leukocyte antigen (BoLA)-DRB3 is a polymorphic gene associated with BLV infectivity and proviral load (PVL). However, the effect of BoLA-DRB3 polymorphism on the infectivity and PVL of milk from BLV-infected dams remains unknown. This study examined milk from 259 BLV-infected dams, including susceptible dams carrying at least one BoLA-DRB3*012:01 or *015:01 allele with high PVL, resistant dams carrying at least one BoLA-DRB3*002:01, *009:02, or *014:01:01 allele with low PVL, and neutral dams carrying other alleles. The detection rate of BLV provirus and PVL were significantly higher in milk from susceptible dams than in that from resistant dams. This result was confirmed in a three-year follow-up study in which milk from susceptible dams showed a higher BLV provirus detection rate over a longer period than that from resistant dams. The visualization of infectivity of milk cells using a luminescence syncytium induction assay showed that the infectious risk of milk from BLV-infected dams was markedly high for susceptible dams compared to resistant ones. This is the first report confirming that BoLA-DRB3 polymorphism affects the PVL and infectivity of milk from BLV-infected dams.

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Section: Blv Infection Rate and Frequencies Of Susceptible Neutral An...mentioning

confidence: 99%

BoLA-DRB3 Polymorphism Controls Proviral Load and Infectivity of Bovine Leukemia Virus (BLV) in Milk

et al. 2022

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“…Next, we measured the PVL of known and novel BLV variants in BLV-infected animals using BLV-CoCoMo-qPCR-2 (RIKEN Genesis, Kanagawa, Japan) [25,[36][37][38]. In total, 142 out of 179 cattle (79.3%) were positive and 37 (20.7%) were negative for BLV PVL (Table 1).…”

Section: Figurementioning

confidence: 99%

Kinetic Study of BLV Infectivity in BLV Susceptible and Resistant Cattle in Japan from 2017 to 2019

et al. 2021

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Bovine leukemia virus (BLV) is the causative agent of enzootic bovine leukosis. Polymorphism in bovine lymphocyte antigen (BoLA)-DRB3 alleles is related to susceptibility to BLV proviral load (PVL), which is a useful index for estimating disease progression and transmission risk. However, whether differential BoLA-DRB3 affects BLV infectivity remains unknown. In a three-year follow-up investigation using a luminescence syncytium induction assay for evaluating BLV infectivity, we visualized and evaluated the kinetics of BLV infectivity in cattle with susceptible, resistant and neutral BoLA-DRB3 alleles which were selected from 179 cattle. Susceptible cattle showed stronger BLV infectivity than both resistant and neutral cattle. The order of intensity of BLV infectivity was as follows: susceptible cattle > neutral cattle > resistant cattle. BLV infectivity showed strong positive correlation with PVL at each testing point. BLV-infected susceptible cattle were found to be at higher risk of horizontal transmission, as they had strong infectivity and high PVL, whereas BLV-infected resistant cattle were low risk of BLV transmission owing to weak BLV infection and low PVL. Thus, this is the first study to demonstrate that the BoLA-DRB3 polymorphism is associated with BLV infection.

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“…К настоящему времени разработано значительное количество ПЦР-методов, направленных на выявление провирусной ДНК в клетках крови животных. Как видно из приведенных выше данных об особенностях организации вирусного генома, наиболее привлекательными для диагностики являются 5'-длинные концевые повторы [42,43], гены gag [44], pol [45], env [46] и tax [47,48]. Важно отметить, что количество копий провируса в общем случае остается чрезвычайно малым по сравнению геномом хозяина, из-за чего большое количество тест-систем сконструировано с использованием гнездовой ПЦР.…”

Section: молекулярно-генетическая диагностика вируса Blvunclassified

Structural characteristics of the bovine leukemia virus genome: A mini review

Fomenko

2023

Food systems

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Enzootic bovine leukemia is an infectious disease with a chronic course caused by an RNA‑containing virus of the genus Deltaretrovirus. Despite the implementation of various programs for the elimination of leukemia, the disease is still widespread on the planet and continues to cause significant economic damage. A large proportion of BLV‑infected cattle remain to be asymptomatic carriers of the virus, which complicates diagnosis and contributes to the spread of the disease in the herd. The structure of the BLV genome is generally typical of retroviruses. It consists of genes encoding structural proteins, viral enzymes and regulatory elements flanked on both sides by identical long terminal repeats. The enzyme and structural protein coding genes (gag, pro, pol, and env) play a crucial role in the life cycle of the virus, influencing its infectivity and virion production. The tax and rex regulatory genes regulate viral transcription, export of transcripts from the nucleus to the cytoplasm, and disease progression. The increase in the number of copies of proviral DNA occurs mainly not due to the functioning of the virus reverse transcriptase, but because of clonal reproduction of the affected subpopulations of B‑cells, mainly CD5+ IgM+. This feature provides increased genetic stability of the BLV virus. These properties of the viral genome allow the development of a variety of PCR test systems. The widespread implementation of such systems enables the detection of carriers of the disease at early stages, which should contribute to the effective implementation of national programs to eradicate bovine leukemia.

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A novel real time PCR assay for bovine leukemia virus detection using mixed probes and degenerate primers targeting novel BLV strains

Cited by 3 publications

References 46 publications

BoLA-DRB3 Polymorphism Controls Proviral Load and Infectivity of Bovine Leukemia Virus (BLV) in Milk

BoLA-DRB3 Polymorphism Controls Proviral Load and Infectivity of Bovine Leukemia Virus (BLV) in Milk

Kinetic Study of BLV Infectivity in BLV Susceptible and Resistant Cattle in Japan from 2017 to 2019

Structural characteristics of the bovine leukemia virus genome: A mini review

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