DETECTION OF PLASMODIUM-FALCIPARUM DNA IN A MICROTITRATION PLATE-BASED HYBRIDIZATION TEST

被引:6
|
作者
KORPELA, K
LAAKSONEN, M
KALLIO, A
SODERLUND, H
PETTERSSON, U
KYRONSEPPA, H
RANKI, M
机构
[1] DEPT MED GENET,UPPSALA,SWEDEN
[2] AURORA HOSP,SF-00250 HELSINKI,FINLAND
关键词
PLASMODIUM-FALCIPARUM; MALARIA; DNA HYBRIDIZATION;
D O I
10.1111/j.1574-6968.1992.tb05147.x
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
A rapid DNA-test, depending on the affinity based hybrid collection principle, was developed for the detection of Plasmodium falciparum DNA from clinical specimens. In this method, hybridization takes place in solution and the hybrids are collected onto a solid phase for measurement. Two probes are used, one labelled with an affinity tag (biotin) and the other with a detectable label (P-32). In the present test a single oligonucleotide complementary to a 21-base pair sequence which is highly repeated in the parasite genome served both as capture and detector probe. The test is a 2-h hybridization performed in streptavidin coated microtitration plate wells, onto which the labelled hybrids simultaneously bind. The sensitivity of the assay with a crude erythrocyte lysate specimen was 1.6 x 10(9) repeat units corresponding to about 160 parasites in one mu-l blood. The results allowed quantification of the repeat sequences and thus estimation of the degree of parasitemia in clinical specimens.
引用
收藏
页码:173 / 178
页数:6
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