A novel recombinase polymerase amplification (RPA) assay for the rapid isothermal detection of Neospora caninum in aborted bovine fetuses.
作者： Ai-Ling Tian , Hany M. Elsheikha , Dong-Hui Zhou , Yao-Dong Wu , Mu-Xin Chen
DOI: 10.1016/J.VETPAR.2018.06.004
关键词:
摘要: The development of a method to rapidly diagnose Neospora caninum infection is highly desirable. Recombinase polymerase amplification (RPA), combined with lateral flow (LF) strips, novel approach amplify and visualize DNA. We have developed prototype LF-RPA assay, using primers probe that targeted specific sequence in the N. NC-5 gene. caninum-specific assay was first tested on purified DNA from oocysts amplified detectable levels 10 min, at constant temperature without need for an expensive thermocycler. designed RPA displayed 100% specificity detecting any cross-reaction nine related protozoan spp. (eg Toxoplasma gondii, Sarcocystis gigantean, zuoi, Hammondia hammondi, heydorni, Eimeria cylindrica, Plasmodium falciparum, Theileria annulata Babesia bigemina). Although, detected amounts as low 50 fg DNA, it nearly 5-fold less sensitive than previously published qPCR nested PCR assays. diagnostic performance detection aborted bovine fetal tissue samples, compared results those obtained PCR. Out 75 samples examined, 18 (24%) 17 (22.6%) positive PCR, respectively. Our indicate suitable rapid reliable caninum.
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