PCR-based quantitation of Cryptosporidium parvum in municipal water samples

被引：18

作者：

Chung, E

Aldom, JE

Carreno, RA

Chagla, AH

Kostrzynska, M

Lee, H ^{[1
]}

Palmateer, G

Trevors, JT

Unger, S

Xu, R

De Grandis, SA

机构：

[1] Univ Guelph, Dept Environm Biol, Guelph, ON N1G 2W1, Canada

[2] Univ Guelph, Lab Serv Div, Guelph, ON N1H 8J7, Canada

[3] Ontario Minist Hlth, Reg Publ Hlth Lab, London, ON N6A 1A4, Canada

[4] GAP Environmicrobial Serv Inc, London, ON N6E 1P5, Canada

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 1999年 / 38卷 / 1-2期

基金：

加拿大自然科学与工程研究理事会;

关键词：

Cryptosporidium parvum; detection; Digene SHARP Signal System (TM); municipal water; oocyst;

D O I：

10.1016/S0167-7012(99)00087-1

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A PCR method for the quantitation of Cryptosporidium parvum oocysts in municipal drinking water samples was investigated. Quantitative PCR uses an internal standard (IS) template with unknown target numbers to compare to standards of known concentrations in a standard curve. The IS template was amplified using the same primers used to amplify a portion of a 358 bp gene fragment that encodes a repetitive oocyst wall protein in C. parvum. Municipal water samples spiked with known numbers of C. parvum oocysts were tested by quantitative PCR using the IS and the Digene SHARP Signal(TM) System Assay for PCR product detection. The absorbance readings for target DNA and IS templates versus the number of molecules of the target DNA were plotted to generate standard curves for estimating oocyst numbers. The method allowed the quantitation of oocysts from log 3 to log 5 spiked into municipal water samples. (C) 1999 Elsevier Science B.V. All rights reserved.

引用

页码：119 / 130

页数：12

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