Comparison of crossmatch results obtained by ELISA, flow cytometry, and conventional methodologies.

The complement-dependent lymphocytotoxicity crossmatch (CXM) is the presently accepted standard for detection of donor-reactive alloantibodies in transplant patients. However, the newer flow cytometric (FXM) and ELISA (EXM) crossmatch technologies are increasingly used as substitutes for the CXM. We have compared the sensitivity and reproducibility of FXM vs. EXM and, in general, find them to be quite similar. However, when we compared the agreement of FXM vs. EXM in 112 donor/recipient combinations, we found that they identified different subsets of donor-specific alloantibodies in about 35% of the tests. When compared to the standard CXM method, the EXM correlated much better than did the FXM, yielding a much lower rate of false positive (2.5% vs. 8%) and false negative (7% vs. 18.5%) results. The reduction in time required to obtain a result (3 h) and the cost of materials ($25/test) was identical for the EXM and FXM. We conclude that the ELISA method for crossmatching has advantages over the flow cytometric method as a substitute for the present standard complement-dependent lymphocytotoxicity method.     

登革病毒悬液芯片分型检测方法的建立

目的 建立一种基于悬液芯片的登革病毒(dengue virus,DV)检测方法,可对四种血清型登革病毒进行快速检测和鉴定.方法 依据GenBank上4种病毒的基因序列信息,设计并合成相关引物及探针序列.抽提病毒RNA,经反转录后对目的基因进行PCR扩增,产物与核酸探针微球组杂交后于Bio-PlexTM 200系统检测荧光信号值.结果 DV1的悬液芯片检测敏感性约9 DNA拷贝,DV2、DV3、DV4的悬液芯片检测敏感性约90 DNA拷贝.进而将本方法用于检测15份临床标本,其检测结果与分型荧光RT-PCR一致.结论 建立了可同时检测四种血清型登革病毒的悬液芯片检测方法,为快速筛查和鉴定登革病毒提供了新的手段.     

Novel multiplex oligonucleotide-conjugated bead suspension array for rapid identification of enterovirus 71 subgenogroups

A high-throughput multiplex bead suspension array was developed for the rapid subgenogrouping of EV71 strains, based on single nucleotide polymorphisms observed within the VP1 region with a high sensitivity as low as 1 PFU. Of 33 viral isolates and 55 clinical samples, all EV71 strains were successfully detected and correctly subgenogrouped.