title: Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces.
abstract: Cryptosporidium is a widespread protozoan parasite that infects a large number of vertebrate animals, resulting in varying degrees of diarrhea or even death. As dairy cattle feces is an important source of Cryptosporidium spp. infection, development of a handy and accurate detection method via its oocysts in dairy cattle feces would be interesting and necessary. We herein developed a quick detecting method using recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip to detect DNA of Cryptosporidium oocysts in dairy cattle feces. The DNA was released by boiled water with 0.1 % N-lauroylsarcosine sodium salt (LSS). The established method was proven to be of higher sensitivity than normal polymerase chain reaction (PCR) amplification with the lowest detection of 0.5 oocyst per reaction, and specificity with no cross reactivity to other common protozoan species in the intestine of dairy cattle. The diagnostic method established herein is simple, rapid, and cost-effective, and has potential for further development as a diagnostic kit for the diagnosis of cryptosporidiosis of dairy cattle.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.
authors.forename: Yao-Dong
authors.initials: YD
authors.lastname: Wu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.
authors.forename: Dong-Hui
authors.initials: DH
authors.lastname: Zhou
authors.affiliation: College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan Province, 450002, People's Republic of China.
authors.forename: Long-Xian
authors.initials: LX
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.|College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province, 130118, People's Republic of China.
authors.forename: Wen-Bin
authors.initials: WB
authors.lastname: Zheng
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.|College of Animal Science and Technology, Jilin Agricultural University, Changchun, Jilin Province, 130118, People's Republic of China.
authors.forename: Jian-Gang
authors.initials: JG
authors.lastname: Ma
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China.
authors.forename: Xing-Quan
authors.initials: XQ
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, 730046, People's Republic of China. xuminjun@scau.edu.cn.|College of Animal Science, South China Agricultural University, Guangzhou, Guangdong Province, 510642, People's Republic of China. xuminjun@scau.edu.cn.
authors.forename: Min-Jun
authors.initials: MJ
authors.lastname: Xu
mesh_terms: D000818:Animals; D002417:Cattle; D002418:Cattle Diseases / Q000175:diagnosis* / Q000469:parasitology; D003457:Cryptosporidiosis / Q000175:diagnosis* / Q000469:parasitology; D003458:Cryptosporidium / Q000235:genetics* / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000235:genetics*; D005243:Feces / Q000469:parasitology*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D033661:Oocysts / Q000166:cytology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012521:Sarcosine / Q000031:analogs & derivatives; D012680:Sensitivity and Specificity
chemical_list: D016054:DNA, Protozoan; C025231:sarkosyl; D012521:Sarcosine
keywords: Cryptosporidium spp.; Dairy cattle; Lateral flow strip; N-lauroylsarcosine sodium salt; Recombinase polymerase amplification (RPA)