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Molecular diagnosis of lyssaviruses and sequence comparison of Australian bat lyssavirus samples.
Aust Vet J. 2006 Jul; 84(7):225-30.AV

Abstract

OBJECTIVE

To evaluate and implement molecular diagnostic tests for the detection of lyssaviruses in Australia.

DESIGN

A published hemi-nested reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of all lyssavirus genotypes was modified to a fully nested RT-PCR format and compared with the original assay. TaqMan assays for the detection of Australian bat lyssavirus (ABLV) were compared with both the nested and hemi-nested RT-PCR assays. The sequences of RT-PCR products were determined to assess sequence variations of the target region (nucleocapsid gene) in samples of ABLV originating from different regions.

RESULTS

The nested RT-PCR assay was highly analytically specific, and at least as analytically sensitive as the hemi-nested assay. The TaqMan assays were highly analytically specific and more analytically sensitive than either RT-PCR assay, with a detection level of approximately 10 genome equivalents per microl. Sequence of the first 544 nucleotides of the nucleocapsid protein coding sequence was obtained from all samples of ABLV received at Australian Animal Health Laboratory during the study period.

CONCLUSION

The nested RT-PCR provided a means for molecular diagnosis of all tested genotypes of lyssavirus including classical rabies virus and Australian bat lyssavirus. The published TaqMan assay proved to be superior to the RT-PCR assays for the detection of ABLV in terms of analytical sensitivity. The TaqMan assay would also be faster and cross contamination is less likely. Nucleotide sequence analyses of samples of ABLV from a wide geographical range in Australia demonstrated the conserved nature of this region of the genome and therefore the suitability of this region for molecular diagnosis.

Authors+Show Affiliations

CSIRO Livestock Industries, Australian Animal Health Laboratory, Private bag 24, Geelong, Victoria 3220.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

16879123

Citation

Foord, A J., et al. "Molecular Diagnosis of Lyssaviruses and Sequence Comparison of Australian Bat Lyssavirus Samples." Australian Veterinary Journal, vol. 84, no. 7, 2006, pp. 225-30.
Foord AJ, Heine HG, Pritchard LI, et al. Molecular diagnosis of lyssaviruses and sequence comparison of Australian bat lyssavirus samples. Aust Vet J. 2006;84(7):225-30.
Foord, A. J., Heine, H. G., Pritchard, L. I., Lunt, R. A., Newberry, K. M., Rootes, C. L., & Boyle, D. B. (2006). Molecular diagnosis of lyssaviruses and sequence comparison of Australian bat lyssavirus samples. Australian Veterinary Journal, 84(7), 225-30.
Foord AJ, et al. Molecular Diagnosis of Lyssaviruses and Sequence Comparison of Australian Bat Lyssavirus Samples. Aust Vet J. 2006;84(7):225-30. PubMed PMID: 16879123.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Molecular diagnosis of lyssaviruses and sequence comparison of Australian bat lyssavirus samples. AU - Foord,A J, AU - Heine,H G, AU - Pritchard,L I, AU - Lunt,R A, AU - Newberry,K M, AU - Rootes,C L, AU - Boyle,D B, PY - 2006/8/2/pubmed PY - 2006/10/25/medline PY - 2006/8/2/entrez SP - 225 EP - 30 JF - Australian veterinary journal JO - Aust Vet J VL - 84 IS - 7 N2 - OBJECTIVE: To evaluate and implement molecular diagnostic tests for the detection of lyssaviruses in Australia. DESIGN: A published hemi-nested reverse transcriptase polymerase chain reaction (RT-PCR) for the detection of all lyssavirus genotypes was modified to a fully nested RT-PCR format and compared with the original assay. TaqMan assays for the detection of Australian bat lyssavirus (ABLV) were compared with both the nested and hemi-nested RT-PCR assays. The sequences of RT-PCR products were determined to assess sequence variations of the target region (nucleocapsid gene) in samples of ABLV originating from different regions. RESULTS: The nested RT-PCR assay was highly analytically specific, and at least as analytically sensitive as the hemi-nested assay. The TaqMan assays were highly analytically specific and more analytically sensitive than either RT-PCR assay, with a detection level of approximately 10 genome equivalents per microl. Sequence of the first 544 nucleotides of the nucleocapsid protein coding sequence was obtained from all samples of ABLV received at Australian Animal Health Laboratory during the study period. CONCLUSION: The nested RT-PCR provided a means for molecular diagnosis of all tested genotypes of lyssavirus including classical rabies virus and Australian bat lyssavirus. The published TaqMan assay proved to be superior to the RT-PCR assays for the detection of ABLV in terms of analytical sensitivity. The TaqMan assay would also be faster and cross contamination is less likely. Nucleotide sequence analyses of samples of ABLV from a wide geographical range in Australia demonstrated the conserved nature of this region of the genome and therefore the suitability of this region for molecular diagnosis. SN - 0005-0423 UR - https://www.unboundmedicine.com/medline/citation/16879123/Molecular_diagnosis_of_lyssaviruses_and_sequence_comparison_of_Australian_bat_lyssavirus_samples_ L2 - https://onlinelibrary.wiley.com/resolve/openurl?genre=article&sid=nlm:pubmed&issn=0005-0423&date=2006&volume=84&issue=7&spage=225 DB - PRIME DP - Unbound Medicine ER -