Detection and differentiation of Schmallenberg, Akabane and Aino viruses by one-step multiplex reverse-transcriptase quantitative PCR assay.
BACKGROUND
Schmallenberg virus (SBV), Akabane virus (AKAV) and Aino virus (AINV) are members of the Simbu serogroup within the genus Orthobunyavirus, family Bunyaviridae, which can cause reproductive disorders including abortion, stillbirth and congenital malformation in ruminants. Because, the clinical signs are similar, confirmatory diagnosis requires viral detection to differentiate infection between these three viruses.


METHODS
In this study, a one-step multiplex reverse-transcriptase quantitative PCR (one-step mRT-qPCR) was developed for the simultaneous detection and differentiation of SBV, AKAV and AINV.


RESULTS
The detection limit of the one-step mRT-qPCR for SBV, AKAV and AINV were 2.4 copies (10 (0.6) TCID 50/ml), 96.2 copies (10 (1.5) TCID 50/ml) and 52.3 copies (10 (1.2) TCID 50/ml), respectively. Various field samples such as bovine serum, bovine whole blood, bovine brain, goat serum and Culicoides were analyzed using the one-step mRT-qPCR and compared with previously published RT-qPCRs. The test results of the field samples were identical for the one-step mRT-qPCR and RT-qPCRs, which showed all samples to be negative for SBV, AKAV and AINV, except for one bovine brain sample (1/123) that was positive for AKAV.


CONCLUSIONS
The one-step mRT-qPCR allows for the simultaneous detection of three viral pathogens (SBV, AKAV and AINV) that cause reproductive failure.
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. leejihye@korea.kr.
Ji-Hye
JH
Lee
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. hyunji3589@korea.kr.
Hyun-Ji
HJ
Seo
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. parkjyyunesy@korea.kr.
Jee-Yong
JY
Park
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. noelove@korea.kr.
Sung-Hee
SH
Kim
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. choys@korea.kr.
Yun Sang
YS
Cho
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. kyjvet@korea.kr.
Yong-Joo
YJ
Kim
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. chois38@korea.kr.
In-Soo
IS
Cho
Animal and Plant Quarantine Agency, Anyang, Gyeonggi-do, 430-824, Republic of Korea. jhy98@korea.kr.
Hye-Young
HY
Jeoung
D000818:Animals; D002044:Bunyaviridae Infections / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D002417:Cattle; D002418:Cattle Diseases / Q000821:virology*; D016851:Orthobunyavirus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012680:Sensitivity and Specificity