title: Rapid detection of Bombyx mori nucleopolyhedrovirus (BmNPV) by loop-mediated isothermal amplification assay combined with a lateral flow dipstick method.
abstract: The Bombyx mori nucleopolyhedrovirus (BmNPV) is a principal pathogen of the domestic silkworm. The disease often breaks out in sericultural countries and due to its high infectivity; it is difficult to control, resulting in heavy economic loss. In order to develop a rapid, sensitive visual detection and simple-to-use novel technology for detection of BmNPV, a loop-mediated isothermal amplification (LAMP) assay combined with a lateral flow dipstick (LFD) method was described. In this study, a set of four primers and a labeled probe were designed specifically to recognize six distinct regions of the BmNPV gp41 gene, and the LAMP for the detection of BmNPV was developed by isothermal amplification at 61 °C for 45 min, followed by hybridization with an FITC-labeled DNA probe for 5 min and detected by LFD within 5 min. The detection limit of LAMP-LFD was 0.2 pg DNA extracted from silkworm infected with BmNPV and was 100 times more sensitive than conventional PCR. No product was generated from silkworm infected with other viruses. Furthermore, we applied the technique to detect BmNPV in the hemolymph and feces at different intervals post infection (pi). In conclusion, the novel LAMP-LFD setup presented here is simple, rapid, reliable, and has the potential for future use in the detection of BmNPV.
authors.affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: College of Animal Science & Technology, Jinling Institute of Technology, Nanjing 210038, China.
authors.forename: Jiege
authors.initials: J
authors.lastname: Wu
authors.affiliation: Key Laboratory of Healthy Freshwater Aquaculture, Ministry of Agriculture, Zhejiang Institute of Freshwater Fisheries, Huzhou 313001, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Lin
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Engineering Technology Research Center of Research and Development of Traditional Chinese Medicine, Lu'an 237012, China.
authors.forename: Naifu
authors.initials: N
authors.lastname: Chen
authors.affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
authors.forename: Shaofei
authors.initials: S
authors.lastname: Yuan
authors.affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
authors.forename: Lina
authors.initials: L
authors.lastname: Ding
authors.affiliation: Institute of Life Sciences, Jiangsu University, Zhenjiang 212013, China.
authors.forename: Li
authors.initials: L
authors.lastname: Gao
authors.affiliation: College of Biological and Pharmaceutical Engineering, West Anhui University, Engineering Technology Research Center of Research and Development of Traditional Chinese Medicine, Lu'an 237012, China. Electronic address: hanbx1978@sina.com.
authors.forename: Bangxing
authors.initials: B
authors.lastname: Hang
mesh_terms: D000818:Animals; D012831:Bombyx / Q000821:virology*; D005243:Feces / Q000821:virology; D006458:Hemolymph / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods* / Q000662:veterinary; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D017924:Nucleopolyhedroviruses / Q000235:genetics / Q000302:isolation & purification*; D015345:Oligonucleotide Probes / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D015345:Oligonucleotide Probes
keywords: Bombyx mori nucleopolyhedrovirus (BmNPV); Lateral flow dipstick (LFD); Loop-mediated isothermal amplification (LAMP); Rapid detection