Closed-tube DNA extraction using a thermostable proteinase is highly sensitive, capable of single parasite detection

被引：14

作者：

Ferrari, B. C. ^{[1
]}

Power, M. L.

Bergquist, P. L.

机构：

[1] Macquarie Univ, Environm Biotechnol CRC, Dept Chem & Biomol Sci, Sydney, NSW 2109, Australia

[2] Macquarie Univ, Biotechnol Res Inst, Sydney, NSW 2109, Australia

[3] Univ Auckland, Sch Med, Dept Microbiol Med & Pathol, Auckland, New Zealand

来源：

BIOTECHNOLOGY LETTERS | 2007年 / 29卷 / 12期

关键词：

DNA extraction; cryptosporidium; giardia; haemonchus; nematoda; proteinase; protozoa;

D O I：

10.1007/s10529-007-9487-2

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：

Current DNA extraction methods for parasites are labour-intensive and usually involve several steps, increasing the potential for cross-contamination. We describe here a closed-tube DNA extraction procedure based upon the use of a thermostable proteinase that enabled sensitive amplification of target loci from parasites from diverse lineages including Apicomplexa, Sarcomastgophora and Nematoda. Moreover, this procedure is not subject to cross-contamination and is readily adaptable to automation.

引用

页码：1831 / 1837

页数：7

共 27 条

[1] A comparison of enumeration techniques for Cryptosporidium parvum oocysts [J].

Bennett, JW ;

Gauci, MR ;

Le Moënic, S ;

Schaefer, FW ;

Lindquist, HDA .

JOURNAL OF PARASITOLOGY, 1999, 85 (06) :1165-1168

[2]

Ferrari BC, 2000, CYTOMETRY, V41, P216, DOI 10.1002/1097-0320(20001101)41:3&lt

[3]

216::AID-CYTO9&gt

[4]

3.0.CO

[5]

2-R

[6] Ribosomal RNA sequencing reveals differences between the genotypes of Giardia isolates recovered from humans and dogs living in the same locality [J].

Hopkins, RM ;

Meloni, BP ;

Groth, DM ;

Wetherall, JD ;

Reynoldson, JA ;

Thompson, RCA .

JOURNAL OF PARASITOLOGY, 1997, 83 (01) :44-51

[7] Development of procedures for direct extraction of Cryptosporidium DNA from water concentrates and for relief of PCR inhibitors [J].

Jiang, JL ;

Alderisio, KA ;

Singh, A ;

Xiao, LH .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2005, 71 (03) :1135-1141

[8] AXENIC CULTURE OF GIARDIA-LAMBLIA IN TYI-S-33 MEDIUM SUPPLEMENTED WITH BILE [J].

KEISTER, DB .

TRANSACTIONS OF THE ROYAL SOCIETY OF TROPICAL MEDICINE AND HYGIENE, 1983, 77 (04) :487-488

[9] Genetic analysis of Cryptosponidium from 2414 humans with diarrhoea in England between 1985 and 2000 [J].

Leoni, F. ;

Amar, C. ;

Nichols, G. ;

Pedraza-Diaz, S. ;

McLauchlin, J. .

JOURNAL OF MEDICAL MICROBIOLOGY, 2006, 55 (06) :703-707

[10] Detection and differentiation of Cryptosporidium parasites that are pathogenic for humans by real-time PCR [J].

Limor, JR ;

Lal, AA ;

Xiao, LH .

JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (07) :2335-2338

← 1 2 3 →