title: Reverse transcription loop-mediated isothermal amplification for sensitive and rapid detection of Korean sacbrood virus.
abstract: Sacbrood virus (SBV) is one of the most serious honeybee viruses. The virus causes failure to pupate and death in both larvae and adult bees. Recently, the Korean sacbrood virus (KSBV) caused great losses in Korean honeybee (Apis cerana) colonies. Although KSBV shows high homology with SBV strains, it has unique motifs and causes different symptoms. Therefore, a simple, sensitive and specific method for detecting KSBV is needed urgently. In this study, a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detecting KSBV using total RNA extracted from honeybees (A. cerana) infected with SBV. The LAMP and the polymerase chain reaction (PCR) methods were then compared for their ability to detect KSBV in clinical samples. The virus was detected in RT-LAMP reactions containing 10(3) copies of pBX-KSBV within 30min, which was comparable to RT-PCR. In addition, the LAMP was able to distinguish between KSBV and other closely-related SBV strains, indicating a high degree of specificity. This simple and sensitive RT-LAMP assay is a useful method for the rapid diagnosis of KSBV infection in honeybees.
authors.affiliation: Animal, Plant and Fisheries Quarantine and Inspection Agency (QIA), 480 Anyangro, Manangu, Anyangsi, Gyeonggido, Republic of Korea. sallala09@hanmail.net
authors.forename: Mi-Sun
authors.initials: MS
authors.lastname: Yoo
authors.forename: Jin-Hyeong
authors.initials: JH
authors.lastname: Noh
authors.forename: Byoung-Su
authors.initials: BS
authors.lastname: Yoon
authors.forename: Kondreddy Eswar
authors.initials: KE
authors.lastname: Reddy
authors.forename: Chang-Hee
authors.initials: CH
authors.lastname: Kweon
authors.forename: Suk-Chan
authors.initials: SC
authors.lastname: Jung
authors.forename: Seung-Won
authors.initials: SW
authors.lastname: Kang
mesh_terms: D000818:Animals; D001516:Bees / Q000821:virology*; D004772:Entomology / Q000379:methods*; D007814:Larva / Q000821:virology; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012328:RNA Viruses / Q000302:isolation & purification*; D056910:Republic of Korea; D012680:Sensitivity and Specificity; D013997:Time Factors; D014773:Virology / Q000379:methods*