Skip to main content

Table 1 Summary of common techniques used for diagnosis of BLV prevalence

From: Epidemiology and genetic diversity of bovine leukemia virus

Diagnostic assay

Sample

Target

Advantages

Disadvantages

References

Type

Assay

Serological test

AGID

Serum

Antibodies (p24, gp51)

Specific, simple, and easy to perform Large scale screening Less expensive

Rapid

Less sensitive and inconclusive Cannot evaluate disease states of infected cattle

Aida et al., 1989 [47]

Wang et al., 1991 [48]

Monti et al., 2005 [49]

Kurdi et al., 1999 [50]

Jimba et al., 2012 [43]

Naif et al., 1990 [55]

ELISA

Serum Milk Bulk milk

Antibodies (p24, gp51)

Specific and sensitive Large scale screening Time saving

False negatives (cattle in early infection phase) False positive (maternally derived antibodies) Cannot evaluate disease states of infected cattle A number of controls and a plate reader required Results require interpretation

Naif et al., 1990 [55]

Burridge et al., 1982 [56]

Schoepf et al., 1997 [53]

Kurdi et al., 1999 [50]

Monti et al., 2005 [49]

Jimba et al., 2012 [43]

Zaghawa et al., 2002 [52]

PHA

Virus particle

BLV glycoprotein

Sensitive Specific detection of BLV Large scale titration Less expensive

Rapid

Affected by pH and temperature Hemagglutination activity reduced by trypsin, potassium periodate, and neuraminidase

Fukai et al., 1999 [51]

RIA

Serum

Antibodies (p24)

Sensitive Able to detect BLV during the early period of infection

Cannot be used for mass screening

Levy et al., 1977 [54]

Nguyen et al., 1993 [57]

Proviral DNA detection

Single PCR; Semi-nested PCR; Nested PCR

Blood PBMC Tumor sample Buffy coat Milk somatic cells

Semen Saliva Nasal secretions

Provirus

Direct, fast, sensitive A variety of samples can be used BLV detection during the early phase of infection or in the presence of colostrum antibodies

Can detect new infections, before the development of antibodies to BLV

Unable to detect BLV when the proviral load is too low

Cross contamination occurs easily Requires specific primers Requires equipment (PCR machine) False negatives in the presence of PCR inhibitory substances in samples Requires internal control Needs confirmatory testing, such as sequencing

Monti et al., 2005 [49]

Kurdi et al., 1999 [50]

Zaghawa et al., 2002 [52]

Tajima et al., 1998 [64]

Tajima et al., 2003 [61]

Real-time PCR

Blood PBMC Tumor sample Buffy coat Milk

Somatic cells Semen Saliva Nasal secretions

Provirus

Direct, fast, sensitive Low risk of contamination A variety of samples can be used Distinguishes EBL from SBL BLV can be detected during the early phase of infection or in the presence of colostrum antibodies Quantitative measurement of proviral load

Requires internal control Requires positive controls of different concentrations Requires specific primers and probes Require equipment (real-time PCR machine) Expensive

Complicated sample preparation procedure

Somura et al., 2014 [68]

Lew et al., 2004 [69]

Jimba et al., 2010 [70]

Jimba et al., 2012 [43]

Tawfeeq et al., 2013 [67]

Brym et al., 2013 [66]

Takeshima et al., 2015 [71]

Direct blood-based PCR

Blood

Provirus

Cost-effective No need for DNA purification Low risk of contamination

Unable to detect BLV when the proviral load is too low

Results in failure if there are mismatches between the PCR primers and BLV sequences Relatively low sensitivity

Nishimori et al., 2016 [72]

Takeshima et al., 2016 [73]

  1. AGID agar gel immunodiffusion, BLV bovine leukemia virus, EBL enzootic bovine leukosis, ELISA enzyme-linked immunosorbent assay, PHA passive hemagglutination assay, RIA radio immunoassay