Specific detection of small ruminant lentiviral nucleic acid sequences located in the proviral long terminal repeat and leader-gag regions using real-time polymerase chain reaction

Brinkhof, J.; Maanen, C. van; Wigger, R.; Lievaart‐Peterson, Karianne; Houwers, D.J.

doi:10.1016/j.jviromet.2007.10.013

Cited by 44 publications

(37 citation statements)

References 37 publications

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“…The PCR method probably gave zero false negatives and had the possibility for diagnosis of CAEV infection by detecting proviral sequences when antibody is absent in circumstances of early infection prior to seroconversion. On the other hand, PCR assay is very useful to prove viral infection in seropositive kids which received maternal antibody from their dams (de Andres et al 2005, Brinkhof et al 2008. When considering the test performance of ELISA, we found satisfactory results as also described by de Andres et al (2005), that the sensitivity values ranged from 92 to 100%.…”

Section: Discussionsupporting

confidence: 64%

“…This finding suggested that in any disease circumstance, ELISA has a possibility of around 95% to detect the truly infected goats and has a possibility of around 83% to indicate the healthy goats. This result depended on the sensitivity that might be caused by the fluctuation of antibody response after infection as well as on the disease prevalence (de Andres et al 2005, Thrusfield 2007, Brinkhof et al 2008 In this study, a relatively high specificity of PCR was expected (Reddy et al 1993, Rimstad et al 1993, Wagter et al 1998, Celer et al 2000, Extramiana et al 2002. This indicated a suitable performance used as a confirmatory test.…”

Section: Discussionmentioning

confidence: 64%

“…It was probably due to viral heterogeneity and low viral loads, which might hamper the application of PCR, resulting in a limitation of this PCR test. The diagnostic value of PCR assay largely depends on the design of the oligonucleotides for priming and of the probes for detecting; therefore, false negative results in this study could be due to mismatch in the primer binding region as also hypothesized with low sensitivity (de Andres et al 2005, Brinkhof et al 2008. The phylogenic study of various strains of circulating viruses and the selection of suitable primer sequences from a relatively conserved region should be emphasized in order to minimize the effects of strain variation and increasing the sensitivity (de Andres et al 2005).…”

mentioning

confidence: 87%

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Diagnosis of Caprine Arthritis Encephalitis Virus infection in dairy goats by ELISA, PCR and Viral Culture

Panneum¹,

Rukkwamsuk²

2017

Polish Journal of Veterinary Sciences

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For preventive and control strategies of Caprine Arthritis Encephalitis Virus (CAEV) infection in dairy goats, performance of the available diagnostic tests was described as one of the most important and necessary aspects. The study aimed at evaluating the diagnostic test performance, including PCR, ELISA and viral culture, for CAEV infection in dairy goats in Thailand. Blood samples of 29 dairy goats from five low-to medium-prevalence herds and one very low-prevalence herd were collected for PCR and ELISA methods. The performance of these two diagnostic methods was evaluated by comparing with cytopathic effects (CPE) in the co-cultivation of CAEV and primary synovial cells. Results indicated that sensitivity, specificity were, respectively, 69.6%, 100%, for PCR; and 95.7%, 83.3% for ELISA. The PCR assay tended to have lower sensitivity and higher specificity than ELISA. When multiple tests were applied, parallel testing provided sensitivity and specificity of 98.7% and 83.3%, while series testing showed sensitivity and specificity of 66.6% and 100% respectively. These results indicated that combination of ELISA and PCR provided some advantages and possibly offered optimal methods to detect CAEV-infected goats. Kappa value of the agreement between PCR and ELISA test was 0.34, indicating fair agreement. Regarding the possibility of antigenic variation between CAEV strains used in both PCR and ELISA assays, the actual circulating CAEV strain should be reviewed in order to develop and enhance the diagnostic tests using the CAE viral antigens derived from specific local strains of Thailand.

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Section: Discussionsupporting

confidence: 64%

Section: Discussionmentioning

confidence: 64%

mentioning

confidence: 87%

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Diagnosis of Caprine Arthritis Encephalitis Virus infection in dairy goats by ELISA, PCR and Viral Culture

Panneum¹,

Rukkwamsuk²

2017

Polish Journal of Veterinary Sciences

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“…Thus, the development of qPCR in this study was followed by genotyping of SRLV strains isolated from infected ewes. Using degenerative primers specific for subtype A12, which is typical for Polish isolates of SRLV, the newly developed qPCR showed high specificity and sensitivity, comparable to those noted for other qPCR assays developed for detection of SRLV infection (5,17). There was a positive correlation between the number of lambs which were positive by qPCR and gag nested PCR at 12 weeks after the birth; however, none of the samples was positive when tested by env nested PCR.…”

Section: Discussionmentioning

confidence: 63%

Presence of specific antibodies and proviral DNA of small ruminant lentiviruses in lambs in their first weeks of life

Olech

Kubis

Lipecka

et al. 2014

Bulletin of the Veterinary Institute in Pulawy

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The aim of this study was to investigate the presence of proviral DNA and colostral antibodies in lambs born to and fed by ewes infected with small ruminant lentiviruses (SRLV). It was demonstrated that all 20 lambs tested 24 h after colostrum ingestion were serologically positive with high antibody titres. These gradually decreased with time, and at week 12 all lambs were seronegative. Twenty percent of lambs tested at the 2 nd week postpartum were provirus positive by qPCR as a result of consumption of infected colostrum or in utero infection. When tested at three months of life, 95% of the lambs were provirus positive, probably as a result of horizontal transmission of the virus. Since these animals could play an important role in the early propagation of SRLV to susceptible herdmates, early removal of provirus-positive animals could help to prevent new infections.

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“…The development of fluorescent methods for PCR and also the instruments like real-time which can monitor the amplification process is considered as a significant improvement in molecular biology. qPCR have been extensively used in various branches of life sciences during the past few years, which includes infectious disease detection in humans (Enbom et al, 2001), animals (Brinkhof et al, 2008) and plants (Boonham et al, 2004), Sample size PCR for TYLCV-OM S0 10 -S1 10 + S2 10 + S3 10 + The (+) sign indicates detection of the virus while the (-) sign indicates that the virus was not detected using PCR (Papayiannis et al, 2009) and mutation scanning (Wittwer et al, 2003).…”

Section: Discussionmentioning

confidence: 99%

Real-time qPCR Assay for the TYLCV Titer in Relation to Symptoms-Based Disease Severity Scales

Ammara

Al-Sadi²,

Al-Shihi

et al. 2017

IJAB

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During the last few decades, Tomato yellow leaf curl disease (TYLCD) has resulted in heavy yield losses and increased incidences in tropical and subtropical regions. In Oman, six different species of begomoviruses were found to be associated with TYLCD. However, Tomato yellow leaf curl virus-Oman (TYLCV-OM) was found to be the most widely distributed species in the whole country. A real-time quantitative PCR (qPCR) assay with an internal control [tomato elongation factor 1(EF1)] was developed for TYLCV-OM, based on SYBRGreen chemistry for the rapid detection and quantification of the virus. This assay was carried out on field samples showing symptoms according to The World Vegetable Center (AVRDC) disease severity scales. The test was used to establish a relationship between virus load and phenotypic symptoms. Viral copies of 2.88 × 10 9 were detected in field infected tomato plants showing symptom severity according to AVRDC scale '3' (i.e. severe leaf curling and yellowing of leaves). The copy number of virus decreased with the relative decrease in symptom severity scale. Tomato plants exhibiting scale '1' and '2' have 7.7 × 10 4 and 7.47 × 10 6 viral copies, respectively. Very low copy number of the virus (564) was detected in tomato plants showing symptoms at a severity scale of '0', which were apparently symptomless. However, tomato plants developed by tissue culture in sterilized conditions, which were used as a negative control, did not show the presence of the virus. The developed qPCR assay could detect as low as 18 fg (femto gram) of virus from total nucleic acid equivalent to approximately 30 genomic units. This quantitative assay can be used to determine virus titer, in breeding programs for development of virus resistant plants and for epidemiological surveys to monitor viral populations and their intensities.

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Specific detection of small ruminant lentiviral nucleic acid sequences located in the proviral long terminal repeat and leader-gag regions using real-time polymerase chain reaction

Cited by 44 publications

References 37 publications

Diagnosis of Caprine Arthritis Encephalitis Virus infection in dairy goats by ELISA, PCR and Viral Culture

Diagnosis of Caprine Arthritis Encephalitis Virus infection in dairy goats by ELISA, PCR and Viral Culture

Presence of specific antibodies and proviral DNA of small ruminant lentiviruses in lambs in their first weeks of life

Real-time qPCR Assay for the TYLCV Titer in Relation to Symptoms-Based Disease Severity Scales

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