Evaluation of fluorochromes for flow cytometric detection of Cryptosporidium parvum oocysts labelled by fluorescent in situ hybridization

被引:13
作者
Deere, D
Vesey, G
Ashbolt, N
Davies, KA
Williams, KL
Veal, D [1 ]
机构
[1] Macquarie Univ, Sch Biol Sci, Ctr Analyt Biotechnol, N Ryde, NSW 2109, Australia
[2] Becton Dickinson Immunocytometry Syst, San Jose, CA 95131 USA
[3] Univ New S Wales, Sch Civil Engn, Kensington, NSW, Australia
关键词
D O I
10.1046/j.1472-765X.1998.00446.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Oligonucleotide probes specific to Cryptosporidium parvum (CRY1) were conjugated with a range of fluorochromes. The fluorescence after in situ hybridization (FISH) labelling of oocysts and controls was assessed. The objective was to determine the most suitable conjugate for FISH labelling, followed by analysis with a 488 nm laser flow cytometer. The most promising candidate was fluorcscein isothiocyanate but only when linked to the CRY1 probe via an 18-carbon spacer arm consisting of six ethylene glycol moieties. The use of the spacer increased fluorescent signals fivefold compared with an equivalent probe in which the FITC was linked directly to the 5'-amino group of the DNA.
引用
收藏
页码:352 / 356
页数:5
相关论文
共 25 条