Development of a nested PCR assay to detect equine infectious anemia proviral DNA from peripheral blood of naturally infected horses

Dong, Jing; Zhu, Wei; Cook, Frank R.; Goto, Yoshitaka; Horii, Yoichiro; Haga, Takeshi

doi:10.1007/s00705-012-1406-8

Cited by 26 publications

(34 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…For EIAV DEV the LTR (Dong et al, ) and gag (Cappelli et al, ), PCRs generated two fragments which covered the 3′ end of the 5′ LTR and the first 500 bases of the gag gene, respectively. Region two of the Irish PCRs (Quinlivan et al, ; primer pair 380F and 1530R) produced an 800 base pair fragment of gag.…”

Section: Resultsmentioning

confidence: 99%

“…The first symptomatic case, EIAV DEV , was used to test the possibility of sequencing by primer walking. Initially, published PCRs designed using sequences from various phylogenetic groupings were used to amplify EIAV DEV , four sets of published PCRs were used: the nested LTR PCR based on the LTR‐gag region (Dong et al, ), the nested gag PCR (Cappelli et al, ), the twelve overlapping PCRs designed to cover the genome of the Irish 2006 outbreak strains (Quinlivan et al, ) and finally, a primer set targeting the gag gene based on the Irish, American, a German and a Japanese strain (Boldabaatar et al, ). Together they included primers based on all EIAV phylogenetic groupings until 2012 (Dong et al, ).…”

Section: Methodsmentioning

confidence: 99%

“…When underpinned by sufficiently large datasets, phylogenetics has proved to be a powerful tool for studying the epidemiology of many diseases including lentiviruses such as Human Immunodeficiency Virus. Full genome EIAV sequences are currently available for the American (EIAV WYO ; Cook et al, ), Chinese (EIAV LIA ; Tu et al, ), Irish (EIAV IRE ; Quinlivan et al, ), Japanese (EIAV MIYA ; Dong et al, ) and Italian cases (EIAV ITA ; Cappelli et al, ) only. The majority of publically available EIAV sequences are only partial genomes covering individual genes.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Molecular characterization of equine infectious anaemia virus strains detected in England in 2010 and 2012

Dorey-Robinson

Locker

Steinbach

et al. 2019

Transbound Emerg Dis

View full text Add to dashboard Cite

Equine infectious anaemia virus (EIAV) is a retrovirus with worldwide distribution which is notifiable to the OIE. Despite its importance to the equine industry, most information regarding its biology have been obtained using only two strains (EIAVWYO and EIAVLIA) from the USA and China, respectively. Recently full genome sequences from Ireland, Italy and Japan have been published; however, this is still not representative of the number of EIAV outbreaks experienced globally each year. The limited availability of published sequences makes design of a universal EIAV PCR difficult, hence diagnosis is solely reliant on serology. Accordingly, it is important to further investigate the re‐emerging cases in other areas of the world. Here, we provide information regarding the outbreaks of EIA in England in 2010 and 2012 including the molecular characterization of strains. Full genome was obtained for two symptomatic cases but could not be resolved for the asymptomatic cases. The two British genomes from 2010 (EIAVDEV) and 2012 (EIAVCOR) each represent a new phylogenetic group, each differing genetically from the other available full genome sequences by 21.1%–25.5%. That the majority of new EIAV full genome sequences to be published adds another phylogenetic group indicates that the surface of EIAV global diversity is just being scratched. These data highlight that further work is needed to fully understand EIAV genetic diversity, namely the full genome sequencing of EIAV cases from a variety of locations and time points. This would aid both the use of phylogenetics in parallel with horse tracing as the epidemiological tool of disease tracking and the design of a universally applicable molecular diagnostic method.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Molecular characterization of equine infectious anaemia virus strains detected in England in 2010 and 2012

Dorey-Robinson

Locker

Steinbach

et al. 2019

Transbound Emerg Dis

View full text Add to dashboard Cite

show abstract

“…During the last few years, the detection of EIAV antibodies directed against p26 protein by ELISA and AGID has been described and these tests are commercialized under various formats [21]. Molecular biological methods such as the nested PCR, multiplex PCR, and real time PCR have been developed for diagnosis of EIAV [5,8,11,18,19,23]. While PCR-based methods represent a potentially highly sensitive alternative for the detection of viral nucleic acids, these techniques require extensive nucleotide sequence information for approval of universal diagnostic method.…”

Section: Discussionmentioning

confidence: 99%

“…ELISA also provides valuable tool for large-scale screening tests, monitoring the prevalence as well as assessing risk factors of EIA. Therefore, in many countries, ELISA has been adopted officially for sero-monitoring of EIAV, with mandatory provision that positive reactions in any EIA ELISA be confirmed by AGID testing before regulatory actions are taken [5,11].…”

Section: Discussionmentioning

confidence: 99%

Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus

et al. 2013

View full text Add to dashboard Cite

Equine infectious anemia (EIA)-a retroviral disease caused by equine infectious anemia virus (EIAV)-is a chronic, debilitating disease of horses, mules, and donkeys. EIAV infection has been reported worldwide and is recognized as pathogen of significant economic importance to the horse industry. This disease falls under regulatory control program in many countries including India. Control of EIA is based on identification of inapparent carriers by detection of antibodies to EIAV in serologic tests and ''Stamping Out'' policy. The current internationally accepted test for diagnosis of EIA is the agar gel immune-diffusion test (AGID), which detects antibodies to the major gag gene (p26) product. The objective of this study was to develop recombinant p26 based in-house immunoassays [enzyme linked immunosorbent assays (ELISA), and AGID] for EIA diagnosis. The synthetic p26 gene of EIAV was expressed in Escherichia coli and diagnostic potential of recombinant p26 protein were evaluated in ELISA and AGID on 7,150 and 1,200 equine serum samples, respectively, and compared with commercial standard AGID kit. The relative sensitivity and specificity of the newly developed ELISA were 100 and 98.6 %, respectively. Whereas, relative sensitivity and specificity of the newly developed AGID were in complete agreement in respect to commercial AGID kit. Here, we have reported the validation of an ELISA and AGID on large number of equine serum samples using recombinant p26 protein produced from synthetic gene which does not require handling of pathogenic EIAV. Since the indigenously developed reagents would be economical than commercial diagnostic kit, the rp26 basedimmunoassays could be adopted for the sero-diagnosis and control of EIA in India.

show abstract

Equine Infectious Anemia Virus

Taylor¹

2017

Interpretation of Equine Laboratory Diagnostics

View full text Add to dashboard Cite

Development of a nested PCR assay to detect equine infectious anemia proviral DNA from peripheral blood of naturally infected horses

Cited by 26 publications

References 17 publications

Molecular characterization of equine infectious anaemia virus strains detected in England in 2010 and 2012

Molecular characterization of equine infectious anaemia virus strains detected in England in 2010 and 2012

Development, evaluation, and laboratory validation of immunoassays for the diagnosis of equine infectious anemia (EIA) using recombinant protein produced from a synthetic p26 gene of EIA virus

Equine Infectious Anemia Virus

Contact Info

Product

Resources

About