title: Development and evaluation of a real-time reverse transcription-loop-mediated isothermal amplification assay for rapid serotyping of foot-and-mouth disease virus.
abstract: A one-step, real-time reverse transcription-loop-mediated isothermal amplification assay (RT-LAMP) for rapid detection and serotyping of Indian foot-and-mouth disease virus (FMDV) is described. The RT-LAMP assay was found to be 10(3)-10(5) fold more sensitive in comparison with RT-PCR, with a detection limit ranging from 10(-3) to 10(-5) TCID(50) of virus samples of all three serotypes. The RT-LAMP assay and qRT-PCR could detect 100 percent of clinical samples of three serotypes, whereas the RT-PCR detected 69.7% of type O, 58.1% of type A and 60.0% of Asia 1 samples. The qRT-PCR has the same sensitivity as the RT-LAMP. The assay conditions with absence of cross reactivity within the three serotypes of FMDV and FMDV negative samples were established. The RT-LAMP assay could detect 100% of samples stored in FTA(®) cards. In comparison with the performance of the RT-PCR; the RT-LAMP appears to be more sensitive, rapid and specific, with the potential for use as a point-of-care (POC) test, especially in developing countries. The use of FTA(®) cards for the preservation of RNA samples coupled with the RT-LAMP assay for the identification of serotypes may help in achieving improved FMDV serotype identification both in the field and in the laboratory.
authors.affiliation: Foot-and-Mouth Disease Virus Laboratory, Research and Development Centre, Indian Immunologicals Limited, Gachibowli, Hyderabad, India.
authors.forename: M
authors.initials: M
authors.lastname: Madhanmohan
authors.forename: S B
authors.initials: SB
authors.lastname: Nagendrakumar
authors.forename: Kankipati
authors.initials: K
authors.lastname: Manikumar
authors.forename: S
authors.initials: S
authors.lastname: Yuvaraj
authors.forename: Satya
authors.initials: S
authors.lastname: Parida
authors.forename: V A
authors.initials: VA
authors.lastname: Srinivasan
mesh_terms: D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D005536:Foot-and-Mouth Disease / Q000175:diagnosis* / Q000821:virology; D029544:Foot-and-Mouth Disease Virus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D057230:Limit of Detection; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis* / Q000235:genetics; D012680:Sensitivity and Specificity; D012703:Serotyping
chemical_list: D012367:RNA, Viral