Bej A K, Mahbubani M H, Dicesare J L, Atlas R M
Department of Biology, University of Louisville, Kentucky 40292.
Appl Environ Microbiol. 1991 Dec;57(12):3529-34. doi: 10.1128/aem.57.12.3529-3534.1991.
To detect low levels of microorganisms in environmental samples by using polymerase chain reaction (PCR)-gene probe detection, samples were concentrated by filtration. Fluoropore (Millipore Corp.) filters were compatible with PCR DNA amplification, whereas various other filters including nitrocellulose and cellulose acetate filters inhibited PCR amplification. By concentrating cells on Fluoropore filters and releasing the DNA by freeze-thaw cycling, PCR DNA amplification could be performed without removing the filter. Concentration with Fluoropore FHLP and FGLP filters permitted the detection of single cells of microorganisms in 100-ml samples by PCR-gene probes.
为了通过聚合酶链反应(PCR)-基因探针检测来检测环境样品中的低水平微生物,样品通过过滤进行浓缩。氟孔(密理博公司)滤膜与PCR DNA扩增兼容,而包括硝酸纤维素和醋酸纤维素滤膜在内的各种其他滤膜则会抑制PCR扩增。通过将细胞浓缩在氟孔滤膜上并通过冻融循环释放DNA,可以在不取下滤膜的情况下进行PCR DNA扩增。使用氟孔FHLP和FGLP滤膜进行浓缩,通过PCR-基因探针能够检测100毫升样品中的单个微生物细胞。