J Vet Sci.  2012 Mar;13(1):43-48. 10.4142/jvs.2012.13.1.43.

Comparison of four diagnostic methods for detecting rabies viruses circulating in Korea

• Yang DK
• Shin EK
• Oh YI
• Lee KW
• Lee CS
• Kim SY
• Lee JA
• Song JY

• KMID: 1365002
• DOI: http://doi.org/10.4142/jvs.2012.13.1.43

Abstract

It is essential to rapidly and precisely diagnose rabies. In this study, we evaluated four diagnostic methods, indirect fluorescent antibody test (FAT), virus isolation (VI), reverse transcriptase polymerase chain reaction (RT-PCR), and rapid immunodiagnostic assay (RIDA), to detect rabies in animal brain homogenates. Out of the 110 animal brain samples tested, 20 (18.2%) were positive for rabies according to the FAT. Compared to the FAT, the sensitivities of VI, RT-PCR, and RIDA were 100, 100, and 95%, respectively. The specificities of VI, RT-PCR and RIDA were found to be 100, 100, and 98.9%, respectively. Rabies viruses circulating in Korea were isolated and propagated in murine neuroblastoma (NG108-15) cells with titers ranging from 101.5 to 104.5 TCID50/mL. Although the RIDA findings did not completely coincide with results obtained from FAT, VI, and RT-PCR, RIDA appears to be a fast and reliable assay that can be used to analyze brain samples. In summary, the results from our study showed that VI, RT-PCR, and RIDA can be used as supplementary diagnostic tools for detecting rabies viruses in both laboratory and field settings.

Keyword

immunodiagnostic assay; rabies; RT-PCR; virus isolation

MeSH Terms

Animals
Antigens, Viral/blood
Brain/virology
Fluorescent Antibody Technique, Indirect/*veterinary
Immunoassay/*veterinary
RNA, Viral/genetics/isolation & purification
Rabies/diagnosis/*veterinary/virology
Rabies virus/genetics/*isolation & purification
Republic of Korea
Reverse Transcriptase Polymerase Chain Reaction/*veterinary
Sensitivity and Specificity

Figure

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  Fig. 1 Rabies-specific fluorescence in the cytoplasm of the infected cells. Rabies field samples (KRVB1001 and KRVB1004) obtained from animals suspected of having rabies and a positive sample [Evelyn-Rokitnicki-Abelseth (ERA) strain] were used to inoculate NG108-15 cells. The cells were then stained with anti-mouse RABV nucleoprotein (N) monoclonal antibody. Uninfected NG108-15 cells stained with the anti-mouse RV N monoclonal antibody were used as a negative control and did not produce any florescence. A: KRVB1001, B: KRVB1004, C: normal NG108-15 cells, D: positive control (ERA strain). × 400.

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  Fig. 2 Result of RT-PCR using primer sets which amplified the nucleoprotein gene of rabies virus. M: 100 bp DNA ladder, Lane 1: KRVR0801, Lane 2: KRVC0802, Lane 3: KRVR0803, Lane 4: KRVR0804, Lane 5: KRVR0901, Lane 6: KRVB0902, Lane 7: KRVB0903, Lane 8: KRVB0904, Lane 9: KRVB0905, Lane 10: KRVR0906, Lane 11: KRVB0907, Lane 12: negative control, Lane 13: positive control, Lane 14: KRVB0908, Lane 15: KRVB0909, Lane 16: KRVB0910, Lane 17: KRVB1001, Lane 18: KRVB1002, Lane 19: KRVC1003, Lane 20: KRVB1004, Lane 21: KRVC1005, Lane 22: KRVB1006, Lane 23: negative control, Lane 24: positive control.

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  Fig. 3 Result of the RIDA after applying the samples (ERA strain, KRVB0904, KRVC1003, and KRVB1004 isolates).

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