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Brief Communication

Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum

Jae-Ran Yu1, Soo-Ung Lee1, Woo-Yoon Park2
The Korean Journal of Parasitology 2009;47(3):293-297.
Published online: August 28, 2009

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Corresponding author (maria205@kku.ac.kr)
• Received: February 24, 2009   • Revised: April 22, 2009   • Accepted: May 16, 2009

Copyright © 2009 by The Korean Society for Parasitology

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Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum
Korean J Parasitol. 2009;47(3):293-297.   Published online August 28, 2009
DOI: https://doi.org/10.3347/kjp.2009.47.3.293
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Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum
Korean J Parasitol. 2009;47(3):293-297.   Published online August 28, 2009
DOI: https://doi.org/10.3347/kjp.2009.47.3.293
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Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum
Image Image
Fig. 1 Position of the primer pair specific to Cryptosporidium parvum gene for PCR and nested PCR. Target genes of C. parvum for PCR were found at GenBank accession no. (COWP, Z22537; SSU rRNA, AF093489; SB012, AF161076).
Fig. 2 Amplification by PCR and nested PCR of Cryptosporidium parvum DNA extracted from purified C. parvum oocysts. The upper gel shows amplification by PCR, and the lower gel shows amplification by nested PCR using amplified PCR products as template DNA. M, 100 bp DNA ladder marker (Promega); (A) COWP gene; (B) COWP-1 gene; (C) SSU rRNA gene; (D) SB012 (random amplified polymorphic DNA).

Fig. 1

Fig. 2

Comparative Sensitivity of PCR Primer Sets for Detection of Cryptosporidium parvum
Target gene Primer Sequences Expected product size (bp) Annealing temperature (℃) Reference (s) COWP Cry-15 5′-GTA GAT AAT GGA AGA GAT TGT G-3′ 550 52 [7,9] Cry-9 5′-GGA CTG AAA TAC AGG CAT TAT CTT G-3′ COWP cowpnest-F1 5′-TGT GTT CAA TCA GAC ACA GC-3′ 311 60 This study cowpnest-R2 5′-TCT GTA TAT CCT GGT GGG C-3′ COWP BcowpF 5′-ACC GCT TCT CAA CAA CCA TCT TGT CCT C-3′ 769 55 [8] BcowpR 5′-CGC ACC TGT TCC CAC TCA ATG TAA ACC C-3′ SSU rRNA rRNA F 5′-TTC TAG AGC TAA TAC ATG CG-3′ 1,325 55 [6] rRNA R 5′-CCC TAA TCC TTC GAA ACA GGA-3′ SSU rRNA nest rRNA F 5′-GGA AGG GTT GTA TTT ATT AGA TAA AG-3′ 826 55 [6] nest rRNA R 5′-AAG GAG TAA GGA ACA ACC TCC A-3′ Random amplified polymorphic DNA SB012F 5′-CTC CGT TCG ATG ATG CAG ATG-3′ 433 51 [5] SB012R 5′-CGG CCC CTG TAG AAA TAA GTC A-3′ No. of C. parvum oocysts COWP PCR COWP nested PCR BCOWPa PCR BCOWPb nested PCR SSU rRNA PCR SSU rRNA nested PCR SB012 PCR 105 +c + + + + + + 104 + + + + + + + 103 -d + + + - + - 102 - + - + - + - 101 - + - - - + - 100 - + - - - - -
Table 1. Primer sets for PCR and nested PCR specific to C. parvum genes

COWP, Cryptosporidium oocyst wall protein.

Table 2. Differences of sensitivity among various primer sets for PCR or nested PCR of C. parvum

PCR performed with the primers, BcowpF and BcowpR;

Nested PCR performed with the primers, Cry-15 and Cry-19, after BCOWP PCR;

+, DNA band was shown by ethidium bromide stained-agarose gel electrophoresis;

-, No DNA band visualized by ethidium bromide stained-agarose gel electrophoresis.

COWP, Cryptosporidium oocyst wall protein.

Table 1.

Table 2.

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