Simultaneous detection of clostridium perfringens and clostridium colinum by duplex-polymerase chain reaction

D. A. Roussan; Ibrahem A. Shaheen; W. S. Totanji; G. Y. Khawaldeh; R. H. Al-Rifai

doi:10.3382/ps.2012-02368

Simultaneous detection of clostridium perfringens and clostridium colinum by duplex-polymerase chain reaction

D. A. Roussan, Ibrahem A. Shaheen, W. S. Totanji, G. Y. Khawaldeh, R. H. Al-Rifai

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

In this study, we provide a protocol for detection of Clostridium colinum and Clostridium perfringens by the single-tube duplex-PCR (dPCR) test for simultaneous and specific detection of both bacteria from pure cultures and fecal samples spiked with these pathogens. Specific primers for each pathogen were selected that amplified products of predicted sizes from bacteria in the dPCR as well as in the single-tube PCR (sPCR) assays. The sensitivity and specificity of dPCR assay were compared with those of the sPCR. No product amplification was obtained with DNA from reference strains belonging to the genus Clostridium (except C. colinum and C. perfringens) and isolates belonging to another genera using these primer sets. The dPCR assay was as sensitive as the sPCR assay because bacterial detection limits were similar in both assays. The detection limits of sPCR and dPCR in bacterial suspension were 20 and 25 cfu/mL for C. colinum and C. perfrin-gens, respectively. Meanwhile, in the presence of feces the sensitivity of both assays decreased to a detection limit of 1.25 × 10⁴ and 1.94 × 10⁴ cfu/g of feces for C. colinum and C. perfringens, respectively. In summary, dPCR assay holds potential to be an economical and rapid diagnostic method for the simultaneous detection of C. colinum and C. perfringens in pure cultures and could be used to screen fecal samples for the presence of these pathogens.

Original language	English
Pages (from-to)	3080-3085
Number of pages	6
Journal	Poultry science
Volume	91
Issue number	12
DOIs	https://doi.org/10.3382/ps.2012-02368
Publication status	Published - Dec 2012
Externally published	Yes

Keywords

Clostridium colinum
Clostridium perfringens
Duplex PCR
Single PCR

ASJC Scopus subject areas

Animal Science and Zoology

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10.3382/ps.2012-02368

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title = "Simultaneous detection of clostridium perfringens and clostridium colinum by duplex-polymerase chain reaction",

abstract = "In this study, we provide a protocol for detection of Clostridium colinum and Clostridium perfringens by the single-tube duplex-PCR (dPCR) test for simultaneous and specific detection of both bacteria from pure cultures and fecal samples spiked with these pathogens. Specific primers for each pathogen were selected that amplified products of predicted sizes from bacteria in the dPCR as well as in the single-tube PCR (sPCR) assays. The sensitivity and specificity of dPCR assay were compared with those of the sPCR. No product amplification was obtained with DNA from reference strains belonging to the genus Clostridium (except C. colinum and C. perfringens) and isolates belonging to another genera using these primer sets. The dPCR assay was as sensitive as the sPCR assay because bacterial detection limits were similar in both assays. The detection limits of sPCR and dPCR in bacterial suspension were 20 and 25 cfu/mL for C. colinum and C. perfrin-gens, respectively. Meanwhile, in the presence of feces the sensitivity of both assays decreased to a detection limit of 1.25 × 104 and 1.94 × 104 cfu/g of feces for C. colinum and C. perfringens, respectively. In summary, dPCR assay holds potential to be an economical and rapid diagnostic method for the simultaneous detection of C. colinum and C. perfringens in pure cultures and could be used to screen fecal samples for the presence of these pathogens.",

keywords = "Clostridium colinum, Clostridium perfringens, Duplex PCR, Single PCR",

author = "Roussan, {D. A.} and Shaheen, {Ibrahem A.} and Totanji, {W. S.} and Khawaldeh, {G. Y.} and Al-Rifai, {R. H.}",

note = "Funding Information: This study was funded by Provimi Jordan (Amman, Jordan). The authors want to thank all members of Provimi Jordan.",

year = "2012",

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doi = "10.3382/ps.2012-02368",

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volume = "91",

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T1 - Simultaneous detection of clostridium perfringens and clostridium colinum by duplex-polymerase chain reaction

AU - Roussan, D. A.

AU - Shaheen, Ibrahem A.

AU - Totanji, W. S.

AU - Khawaldeh, G. Y.

AU - Al-Rifai, R. H.

N1 - Funding Information: This study was funded by Provimi Jordan (Amman, Jordan). The authors want to thank all members of Provimi Jordan.

PY - 2012/12

Y1 - 2012/12

N2 - In this study, we provide a protocol for detection of Clostridium colinum and Clostridium perfringens by the single-tube duplex-PCR (dPCR) test for simultaneous and specific detection of both bacteria from pure cultures and fecal samples spiked with these pathogens. Specific primers for each pathogen were selected that amplified products of predicted sizes from bacteria in the dPCR as well as in the single-tube PCR (sPCR) assays. The sensitivity and specificity of dPCR assay were compared with those of the sPCR. No product amplification was obtained with DNA from reference strains belonging to the genus Clostridium (except C. colinum and C. perfringens) and isolates belonging to another genera using these primer sets. The dPCR assay was as sensitive as the sPCR assay because bacterial detection limits were similar in both assays. The detection limits of sPCR and dPCR in bacterial suspension were 20 and 25 cfu/mL for C. colinum and C. perfrin-gens, respectively. Meanwhile, in the presence of feces the sensitivity of both assays decreased to a detection limit of 1.25 × 104 and 1.94 × 104 cfu/g of feces for C. colinum and C. perfringens, respectively. In summary, dPCR assay holds potential to be an economical and rapid diagnostic method for the simultaneous detection of C. colinum and C. perfringens in pure cultures and could be used to screen fecal samples for the presence of these pathogens.

AB - In this study, we provide a protocol for detection of Clostridium colinum and Clostridium perfringens by the single-tube duplex-PCR (dPCR) test for simultaneous and specific detection of both bacteria from pure cultures and fecal samples spiked with these pathogens. Specific primers for each pathogen were selected that amplified products of predicted sizes from bacteria in the dPCR as well as in the single-tube PCR (sPCR) assays. The sensitivity and specificity of dPCR assay were compared with those of the sPCR. No product amplification was obtained with DNA from reference strains belonging to the genus Clostridium (except C. colinum and C. perfringens) and isolates belonging to another genera using these primer sets. The dPCR assay was as sensitive as the sPCR assay because bacterial detection limits were similar in both assays. The detection limits of sPCR and dPCR in bacterial suspension were 20 and 25 cfu/mL for C. colinum and C. perfrin-gens, respectively. Meanwhile, in the presence of feces the sensitivity of both assays decreased to a detection limit of 1.25 × 104 and 1.94 × 104 cfu/g of feces for C. colinum and C. perfringens, respectively. In summary, dPCR assay holds potential to be an economical and rapid diagnostic method for the simultaneous detection of C. colinum and C. perfringens in pure cultures and could be used to screen fecal samples for the presence of these pathogens.

KW - Clostridium colinum

KW - Clostridium perfringens

KW - Duplex PCR

KW - Single PCR

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JF - Poultry science

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ER -

Simultaneous detection of clostridium perfringens and clostridium colinum by duplex-polymerase chain reaction

Abstract

Keywords

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