Sensitive and specific detection of bovine immunodeficiency virus and bovine syncytial virus by 5′ Taq nuclease assays with fluorescent 3′ minor groove binder-DNA probes

Lew, Ala E.; Bock, R.E.; Miles, John J.; Cuttell, Leigh; Steer, Penelope A.; Nadin-Davis, Susan A.

doi:10.1016/j.jviromet.2003.10.006

Cited by 21 publications

(18 citation statements)

References 41 publications

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“…Phosphorimager quantification of these hybridization signals revealed that mutants IG4, CRX3 and CRE replicated at lower proviral loads compared to the wild type or the non-attenuated GP30 viruses (Figure 5B). Using real-time PCR performed as described by [26], the proviral loads were estimated to be 270 and 245 copies per 1,000 cells in sheep infected, respectively, with the wild type virus and the GP30 recombinant whereas the levels yielded by the other mutants were significantly reduced (27, 40 and 24 copies / 1,000 cells for IG4, CRX3 and CRE, respectively) (Figure 5B). …”

Section: Resultsmentioning

confidence: 99%

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Section: Resultsmentioning

confidence: 99%

“…Real-time PCR was performed using 6FAM-labeled MGB probes specific for the BLV pol gene and the 18S ribosomal DNA sequences essentially as described in reference [26]. …”

Section: Methodsmentioning

confidence: 99%

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“…Therefore, a more accurate way to monitor BLV infection is PCR [100,101]. BLV-infected cattle can further be classified according to their proviral load (PVL) [102].…”

Section: Preventive and Therapeutic Strategiesmentioning

confidence: 99%

Preventive and Therapeutic Strategies for Bovine Leukemia Virus: Lessons for HTLV

Rodríguez

Florins

Gillet

et al. 2011

Viruses

172

126

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Bovine leukemia virus (BLV) is a retrovirus closely related to the human T-lymphotropic virus type 1 (HTLV-1). BLV is a major animal health problem worldwide causing important economic losses. A series of attempts were developed to reduce prevalence, chiefly by eradication of infected cattle, segregation of BLV-free animals and vaccination. Although having been instrumental in regions such as the EU, these strategies were unsuccessful elsewhere mainly due to economic costs, management restrictions and lack of an efficient vaccine. This review, which summarizes the different attempts previously developed to decrease seroprevalence of BLV, may be informative for management of HTLV-1 infection. We also propose a new approach based on competitive infection with virus deletants aiming at reducing proviral loads.

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“…paratuberculosis (28). Minor groove binder (MGB) probes demonstrate higher specificities and sensitivities than non-MGB probes in 5Ј Taq nuclease assays (30) and thus are highly suited for routine diagnostic applications as demonstrated for the detection of bovine retroviruses (34,35). This study describes the optimization of sampling, transport, and processing protocols for the diagnosis of bovine venereal campylobacteriosis by using a novel 5Ј Taq nuclease PCR assay utilizing a 3Ј TaqMan MGB probe.…”

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Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

McMillen¹,

Fordyce²,

Doogan³

et al. 2006

J Clin Microbiol

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A Campylobacter fetus subsp. venerealis-specific 5 Taq nuclease PCR assay using a 3 minor groove binder-DNA probe (TaqMan MGB) was developed based on a subspecies-specific fragment of unknown identity (S. Hum, K. Quinn, J. Brunner, and S. L. On, Aust. Vet. J. 75:827-831, 1997). The assay specifically detected four C. fetus subsp. venerealis strains with no observed cross-reaction with C. fetus subsp. fetus-related Campylobacter species or other bovine venereal microflora. The 5 Taq nuclease assay detected approximately one single cell compared to 100 and 10 cells in the conventional PCR assay and 2,500 and 25,000 cells from selective culture from inoculated smegma and mucus, respectively. The respective detection limits following the enrichments from smegma and mucus were 5,000 and 50 cells/inoculum for the conventional PCR compared to 500 and 50 cells/inoculum for the 5 Taq nuclease assay. Field sampling confirmed the sensitivity and the specificity of the 5 Taq nuclease assay by detecting an additional 40 bulls that were not detected by culture. Urine-inoculated samples demonstrated comparable detection of C. fetus subsp. venerealis by both culture and the 5 Taq nuclease assay; however, urine was found to be less effective than smegma for bull sampling. Three infected bulls were tested repetitively to compare sampling tools, and the bull rasper proved to be the most suitable, as evidenced by the improved ease of specimen collection and the consistent detection of higher levels of C. fetus subsp. venerealis. The 5 Taq nuclease assay demonstrates a statistically significant association with culture ( 2 ‫؍‬ 29.8; P < 0.001) and significant improvements for the detection of C. fetus subsp. venerealis-infected animals from crude clinical extracts following prolonged transport.

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Sensitive and specific detection of bovine immunodeficiency virus and bovine syncytial virus by 5′ Taq nuclease assays with fluorescent 3′ minor groove binder-DNA probes

Cited by 21 publications

References 41 publications

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Preventive and Therapeutic Strategies for Bovine Leukemia Virus: Lessons for HTLV

Comparison of Culture and a Novel 5′ Taq Nuclease Assay for Direct Detection of Campylobacter fetus subsp. venerealis in Clinical Specimens from Cattle

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