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Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces.
Parasitol Res 2016; 115(9):3551-5PR

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+Show Affiliations

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.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 Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan Province, 450002, People's Republic of China.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.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.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.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.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.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

27174027

Citation

Wu, Yao-Dong, et al. "Recombinase Polymerase Amplification (RPA) Combined With Lateral Flow (LF) Strip for Equipment-free Detection of Cryptosporidium Spp. Oocysts in Dairy Cattle Feces." Parasitology Research, vol. 115, no. 9, 2016, pp. 3551-5.
Wu YD, Zhou DH, Zhang LX, et al. Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces. Parasitol Res. 2016;115(9):3551-5.
Wu, Y. D., Zhou, D. H., Zhang, L. X., Zheng, W. B., Ma, J. G., Wang, M., ... Xu, M. J. (2016). Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces. Parasitology Research, 115(9), pp. 3551-5. doi:10.1007/s00436-016-5120-4.
Wu YD, et al. Recombinase Polymerase Amplification (RPA) Combined With Lateral Flow (LF) Strip for Equipment-free Detection of Cryptosporidium Spp. Oocysts in Dairy Cattle Feces. Parasitol Res. 2016;115(9):3551-5. PubMed PMID: 27174027.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Recombinase polymerase amplification (RPA) combined with lateral flow (LF) strip for equipment-free detection of Cryptosporidium spp. oocysts in dairy cattle feces. AU - Wu,Yao-Dong, AU - Zhou,Dong-Hui, AU - Zhang,Long-Xian, AU - Zheng,Wen-Bin, AU - Ma,Jian-Gang, AU - Wang,Meng, AU - Zhu,Xing-Quan, AU - Xu,Min-Jun, Y1 - 2016/05/13/ PY - 2016/04/15/received PY - 2016/05/04/accepted PY - 2016/5/14/entrez PY - 2016/5/14/pubmed PY - 2017/6/1/medline KW - Cryptosporidium spp. KW - Dairy cattle KW - Lateral flow strip KW - N-lauroylsarcosine sodium salt KW - Recombinase polymerase amplification (RPA) SP - 3551 EP - 5 JF - Parasitology research JO - Parasitol. Res. VL - 115 IS - 9 N2 - 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. SN - 1432-1955 UR - https://www.unboundmedicine.com/medline/citation/27174027/Recombinase_polymerase_amplification__RPA__combined_with_lateral_flow__LF__strip_for_equipment_free_detection_of_Cryptosporidium_spp__oocysts_in_dairy_cattle_feces_ L2 - https://dx.doi.org/10.1007/s00436-016-5120-4 DB - PRIME DP - Unbound Medicine ER -