Establishment of a quantitative real-time PCR for detecting Babesia spp. and the investigation on the prevalence of Babesia spp. in the voluntary blood donors in Mudanjiang city
YANG Yashang;GUO Yanli;LI Hongjun;HUANG Yang;WU Wenjie;TAN Baohua;HE Miao;LIU Yu;Institute of Blood Transfusion,Chinese Academy of Medical Sciences;Sichuan Blood Safety and Blood Substiute,International Science and Technology Cooperation Base;Mudanjang blood center;
Objective To establish a quantitative real-time PCR method for screening a variety of infectious Babesia spp.( Including B. microti,B. divergens,B.venatorum,Babesia.sp EU1,Babesia_sp._XXB/Hangzhou,); two methods including quantitative real-time PCR and indirect immunofluorescence assay( IFA) were used to test the blood samples of the voluntary blood donors in Mudanjiang City,and then assess the risk of transfusion safety. Methods The gene sequences of all species of babesia were downloaded from Genbank and primers were designed based on the 18 s r RNA conserved region. The real-time PCR detection method was established after necessary optimizations.The nucleic acid test was carried out with realtime PCR after mixing samples( 10 parts/pool) of 1971 red blood cell( RBC) samples and then a split detection was performed on the positive pools; the plasma samples of 1000 blood donors were tested by IFA for B. microti Ig G. Results A real-time PCR detection method for Babesia spp. was successfully established and its lowest detected limitation was 9. 69× 102 copies/m L.The results of Babesia spp. nucleic acid with 1971 blood samples were all negative; 13 cases( 1. 3%) were positive for B. microti Ig G detection within 1000 plasma samples.The titer of the 8 cases and the 5 cases were 1: 64 and 1: 128,respectively. Conclusion In this study,a quantitative real-time PCR detection methods screening various infectious human babesia was successfully developed,which can be directly used for screening. The results of IFA detection indicated that there was previous B. microti infection in Mudanjiang area. And the risk of babesia in blood transfusions did exist,which requires further assessment to evaluate the threat.
【CateGory Index】： R531
【CateGory Index】： R531