title: Rapid, simple and direct detection of Meloidogyne hapla from infected root galls using loop-mediated isothermal amplification combined with FTA technology.
abstract: The northern root-knot nematode (Meloidogyne hapla) is a damaging nematode that has caused serious economic losses worldwide. In the present study, a sensitive, simple and rapid method was developed for detection of M. hapla in infested plant roots by combining a Flinders Technology Associates (FTA) card with loop-mediated isothermal amplification (LAMP). The specific primers of LAMP were designed based on the distinction of internal transcribed spacer (ITS) sequences between M. hapla and other Meloidogyne spp. The LAMP assay can detect nematode genomic DNA at concentrations low to 1/200 000, which is 100 times more sensitive than conventional PCR. The LAMP was able to highly specifically distinguish M. hapla from other closely related nematode species. Furthermore, the advantages of the FTA-LAMP assay to detect M. hapla were demonstrated by assaying infected root galls that were artificially inoculated. In addition, M. hapla was successfully detected from six of forty-two field samples using FTA-LAMP technology. This study was the first to provide a simple diagnostic assay for M. hapla using the LAMP assay combined with FTA technology. In conclusion, the new FTA-LAMP assay has the potential for diagnosing infestation in the field and managing the pathogen M. hapla.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Peng
authors.affiliation: Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China.
authors.forename: Haibo
authors.initials: H
authors.lastname: Long
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Wenkun
authors.initials: W
authors.lastname: Huang
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Jiangkuan
authors.initials: J
authors.lastname: Cui
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Lingan
authors.initials: L
authors.lastname: Kong
authors.affiliation: The National Engineering Research Center of Agribiodiversity Applied Technologies, Yunnan Agricultural University, Kunming 650201, China.
authors.forename: Xianqi
authors.initials: X
authors.lastname: Hu
authors.affiliation: Academy of Inspection and Quarantine, Ningbo, Zhejiang, 315012, China.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Gu
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
authors.forename: Deliang
authors.initials: D
authors.lastname: Peng
mesh_terms: D000818:Animals; D021901:DNA, Intergenic; D016054:DNA, Protozoan; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D010935:Plant Diseases / Q000469:parasitology*; D018517:Plant Roots / Q000469:parasitology*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014431:Tylenchoidea / Q000235:genetics*
chemical_list: D021901:DNA, Intergenic; D016054:DNA, Protozoan