对从污水和河水中分离出的单个隐孢子虫卵囊进行基因分型。

Genotyping of single Cryptosporidium oocysts isolated from sewage and river water.

作者信息

Hirata T, Hashimoto A

机构信息

Water Environment Laboratory, School of Environmental Health, Azabu University, 1-17-71 Fuchinobe, Sagamihara, 229-8501 Kanagawa, Japan.

出版信息

Water Sci Technol. 2006;54(3):197-202. doi: 10.2166/wst.2006.469.

DOI:10.2166/wst.2006.469

PMID:17037153

Abstract

The study was designed to genotype individual Cryptosporidium oocysts using an 18S rRNA gene-based semi-nested PCR and direct sequencing procedure. Positive PCR amplification was observed in all single C. parvum HNJ-1 oocyst samples tested. Semi-nested PCR and direct sequencing was applied to Cryptosporidium oocysts isolated from sewage and river water. The procedure could genotype 54% of FITC-stained single oocysts isolated from sewage and 32% from river water. The predominant genotype in both sewage and river water was C. parvum genotype 1, accounting for 33 and 25%, respectively, of all the FITC-stained intact Cryptosporidium oocysts present.

摘要

本研究旨在使用基于18S rRNA基因的半巢式PCR和直接测序程序对单个隐孢子虫卵囊进行基因分型。在所有检测的单个微小隐孢子虫HNJ-1卵囊样本中均观察到阳性PCR扩增。半巢式PCR和直接测序应用于从污水和河水中分离的隐孢子虫卵囊。该程序能够对从污水中分离的54%的异硫氰酸荧光素（FITC）染色单个卵囊以及从河水中分离的32%的卵囊进行基因分型。污水和河水中的主要基因型均为微小隐孢子虫基因型1，分别占所有FITC染色完整隐孢子虫卵囊的33%和25%。