Specific PCR primers for Cryptosporidium parvum with extra high sensitivity

被引：26

作者：

Wu, Z

Nagano, I

Matsuo, A

Uga, S

Kimata, I

Iseki, M

Takahashi, Y

机构：

[1] Gifu Univ, Sch Med, Dept Parasitol, Gifu 5008705, Japan

[2] Kobe Univ, Sch Med, Fac Hlth Sci, Dept Med Technol,Suma Ku, Kobe, Hyogo 6500017, Japan

[3] Osaka City Univ, Sch Med, Dept Med Zool, Abeno Ku, Osaka 5458585, Japan

来源：

MOLECULAR AND CELLULAR PROBES | 2000年 / 14卷 / 01期

关键词：

Cryptosporidium parvum; PCR; primer;

D O I：

10.1006/mcpr.1999.0280

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

One pair of high-sensitive polymerase chain reaction (PCR) primers for Cryptosporidium parvum was constructed based on the sequence of random amplified polymorphic DNA. PCR with this primer pair amplified only the DNA of C. parvum, not the control DNA including Cryptosporidium muris. This primer pair had most advantageous in its sensitivity over the six pairs of primers reported elsewhere. The minimum amount of template DNA required to produce visible bands after gel electrophoresis and ethidium bromide staining was 0.156 pg of C. parvum or just a single oocyst in the PCR tube. (C) 2000 Academic Press.

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收藏

页码：33 / 39

页数：7

相关论文

共 21 条

[1] DETECTION OF CRYPTOSPORIDIUM-PARVUM OOCYSTS IN BOVINE FECES BY MONOCLONAL-ANTIBODY CAPTURE ENZYME-LINKED-IMMUNOSORBENT-ASSAY [J].

ANUSZ, KZ ;

MASON, PH ;

RIGGS, MW ;

PERRYMAN, LE .

JOURNAL OF CLINICAL MICROBIOLOGY, 1990, 28 (12) :2770-2774

[2] THE DEVELOPMENT AND PERFORMANCE OF A SIMPLE, SENSITIVE METHOD FOR THE DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN FECES [J].

BAXBY, D ;

BLUNDELL, N ;

HART, CA .

JOURNAL OF HYGIENE, 1984, 93 (02) :317-&

[3] PCR CLONING AND NUCLEOTIDE-SEQUENCE DETERMINATION OF THE 18S RIBOSOMAL-RNA GENES AND INTERNAL TRANSCRIBED SPACER-1 OF THE PROTOZOAN PARASITES CRYPTOSPORIDIUM-PARVUM AND CRYPTOSPORIDIUM-MURIS [J].

CAI, J ;

COLLINS, MD ;

MCDONALD, V ;

THOMPSON, DE .

BIOCHIMICA ET BIOPHYSICA ACTA, 1992, 1131 (03) :317-320

[4] A new restriction fragment length polymorphism from Cryptosporidium parvum identifies genetically heterogeneous parasite populations and genotypic changes following transmission from bovine to human hosts [J].

Carraway, M ;

Tzipori, S ;

Widmer, G .

INFECTION AND IMMUNITY, 1997, 65 (09) :3958-3960

[5] CRYPTOSPORIDIOSIS [J].

CURRENT, WL ;

GARCIA, LS .

CLINICAL MICROBIOLOGY REVIEWS, 1991, 4 (03) :325-358

[6] DEVELOPMENT OF A PCR PROTOCOL FOR SENSITIVE DETECTION OF CRYPTOSPORIDIUM OOCYSTS IN WATER SAMPLES [J].

JOHNSON, DW ;

PIENIAZEK, NJ ;

GRIFFIN, DW ;

MISENER, L ;

ROSE, JB .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 1995, 61 (11) :3849-3855

[7]

Kaucner C, 1998, APPL ENVIRON MICROB, V64, P1743

[8] The development of diagnostic PCR primers for Cryptosporidium using RAPD-PCR [J].

Morgan, UM ;

OBrien, PA ;

Thompson, RCA .

MOLECULAR AND BIOCHEMICAL PARASITOLOGY, 1996, 77 (01) :103-108

[9] Differentiation between human and animal isolates of Cryptosporidium parvum using rDNA sequencing and direct PCR analysis [J].

Morgan, UM ;

Constantine, CC ;

Forbes, DA ;

Thompson, RCA .

JOURNAL OF PARASITOLOGY, 1997, 83 (05) :825-830

[10] PCR detection of Cryptosporidium:: The way forward? [J].

Morgan, UM ;

Thompson, RCA .

PARASITOLOGY TODAY, 1998, 14 (06) :241-245