title: Development of loop-mediated isothermal amplification for rapid detection of avian leukosis virus subgroup A.
abstract: This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for distinguishing avian leukosis virus (ALV) subgroup A from other subgroups of the virus. On the basis of the results of sequence comparison and the sequence characteristics of ALV subgroups, a LAMP method was designed to target the gp85 segment for detection of ALV-A. Under optimal reaction conditions, ALV-A LAMP produced neither cross-reactions with other major subgroups (including subgroups J, B, C, and E) nor nonspecific reactions with other common avian infectious diseases. A sensitivity test showed that this method can detect 20 copies of proviral nucleic acid sequence within 45 min, which is 100 times more sensitive than the conventional polymerase chain reaction (PCR). This method can detect subgroup A virus rapidly and the results can be assessed based on color changes. The whole reaction process can be performed without opening the lid of the reaction tube, which reduces the possibility of contamination greatly and simplifies the detection process, indicating the considerable potential of this method for in situ application in the future.
authors.affiliation: The Chinese Academy of Agricultural Sciences, Harbin, Heilongjiang 150001, China.
authors.forename: Yongqiang
authors.initials: Y
authors.lastname: Wang
authors.forename: Zhonghui
authors.initials: Z
authors.lastname: Kang
authors.forename: Yulong
authors.initials: Y
authors.lastname: Gao
authors.forename: Liting
authors.initials: L
authors.lastname: Qin
authors.forename: Lei
authors.initials: L
authors.lastname: Chen
authors.forename: Qi
authors.initials: Q
authors.lastname: Wang
authors.forename: Jiukuan
authors.initials: J
authors.lastname: Li
authors.forename: Honglei
authors.initials: H
authors.lastname: Gao
authors.forename: Xiaole
authors.initials: X
authors.lastname: Qi
authors.forename: Huan
authors.initials: H
authors.lastname: Lin
authors.forename: Xiaomei
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D001353:Avian Leukosis / Q000175:diagnosis / Q000821:virology*; D001354:Avian Leukosis Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D002645:Chickens; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D012680:Sensitivity and Specificity; D013696:Temperature; D014773:Virology / Q000379:methods*