Detection of Cpegene in Stool Samples of Food Poisoning Patients in Mosul/Iraq
Keywords:
Clostridium perfringens, Food poisoning, Nested PCR, Cpe geneAbstract
In this study, 27 stool samples collected from food poisoning patients in Mosul city were analyzed by both bacteriological standard methods and molecular methods. The bacteriological methods used for enumeration of C. perfringens while the nested PCR technique was used for detection cpe gene directly in stool samples in a method consists of a combination of nested PCR reaction with enrichment of the sample. Two pairs of oligonucleotide primers were used: the first primer pair amplifies a 425-bp fragment and the second pair amplifiesa199-bp within the 425-bp. Results showed that the number of C. perfringens were less than 103cell/g gave a 199-bp amplified fragment which indicated that those samples contain cpe gene. When we compared nested PCR result with the number of C. perfringens,7 out of 9 samples(77. 8%) which have greater than 103of C. perfringens per g were positive for cpe gene and 8 out of 18 samples(44. 4%) which have less than 103 of C. perfringens per g also gave cpe gene positive result.
References
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Asha, N.J., and Wilcox,M.H. (2002).Laboratory diagnosis of Clostridiumperfringens antibiotic-associated diarrhea. Journal ofMedicalMicrobiology, 51,891-894.
Berry, P.R.,Rodhouse, J.C.,Hughes, S.,Batholomew,B.A,and Gilbert, R.J. (1988).Evaluation of ELISA,RPLA and Vero cell assay for detecting Clostridiumperfringens enterotoxin in feacal specimens. JournalofClinicalPathologyl,41,458-461.
Eisgrüber, H., and Hauner, G. (2001).Minead beef heart associated with a Clostridiumperfringens food poisoning in a Munich old people’s home. ArchLebensmihelhyg, 52,63-66.
Fach,P.,andPopoff,M.R.(1997).Detection of enerotoxigenicClostridiumperfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test. Applied and EnvironmentalMicrobiology, 63,4232-4236.
Gumerlock, P.H., Tang, Y.J., Weiss,J.B.,andSliva,J.Jr. (1993).Specific detection of toxigenic strains of Clostridiumdefficilein stool specimens. JournalofClinicalMicrobiology, 31,507-511.
Jay, J.M. (2000).Modern Food Microbiology(6thed.).Gaithersburg:AspenPuplishers.
Lahti, P., Heikinheimo, A., Johansson,T.,andKorkeala, H. (2008).Clostridiumperfringens type A strains carrying a plasmid-borne enterotoxin gene (genotype IS1151-cpe or IS1470-like-cpe) as a common cause of food poisoning .JournalofClinicalMicrobiology, 46,371-373.
Le Marc, Y., Plowman, J., Aldus, C.F., Munoz-Cuevas, M., Baranyi,J.,and Peck, M.W. (2008).Modelling the growth of Clostridiumperfringens during the cooling of bulk meat.InternationalJournalofFoodMicrobiology ,128,41-50.
Lukinmaa, S., Takkunen,E.,andSiitonen, A. (2002). Molecular epidemiology of Clostridiumperfringens related to food-borne outbreaks of disease in Finland from 1984 to 1999.AppliedandEnvironmentalMicrobiology,68,3744-3749.
Matches, J.R.,Liston,T.,andCurran, D. (1974).Clostridiumperfringens in the environment. AppliedMicrobiology, 28,655-660.
McClane, B.A. (2005).Clostridialenterotoxins. In: P .Dure(Eds),Handbook on Clostridia (pp. 385-406). BocaRaton:CRC Press.
McClane, B.A., Uzal,F.A., Miyakawa,M.F.,Lyerly,D.,and Wilkins, T. (2006).The enterotoxic Clostridia. In:M .Dworkin,S.Falkow,E. Rosen-burg,K.H.Schlcifer&E .Stackebrandt(Eds),The Prokaryotes,vol4(pp 698-752). New York:Springer-Verlag.
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McClane,B.A.,andStrouse, R.J.(1984).Rapid detection of Clostridiumperfringens type A enterotoxin by enzyme-linked immunosorbent assay. JournalofClinicalMicrobiology, 19,112-115.
Miwa, N., Nishina, T. , Kubo,S. ,and Fujikura, K. (1996).Nested polymerase chain reaction for detection oflow levels of enterotoxigenicClosridiumperfringens in animal feces and meat . Journal ofVeterinaryMedical Science,58,197-203.
Monma, C.,Yanagawa, Y., Kusunoki, J., Kai, A.,Shingaki, M., Obata, H., Itoh, T., Ohta, K., and Kudoh, Y. (1994). Serotyping and enterotoxin production of heat-resistant spore-forming Clostridiumperfringens isolated from human feces.Annual Report ofTokyoMetropolitanResearchLaboratory ofPuplicHealth,45,11-15.
Nakamura,H., Ogasawara,J. , Monna, C., Suzuki,A.H., Kai,A.,Haruki,K.,andNishikawa, Y. (2003).Usefulness of a combination of pulsed-field gel electrophoresis and enrichment culture in laboratory invetigation of foodborne outbreak due to Clostridiumperfringens. DiagnosticMicrobiology andInfectiousDisease,47,471-475.
Petit, L.,Gilbert,M.,andPopoff, M.R. (1999) .Clostridiumperfringens: toxinotype and genotype. Trends in Microbiology, 7,104-110.
Rood, J.I. (1998).Virulance genes of Clostridiumperfringens. AnnualReviewofMicrobiology,52,333-360.
Smedley, J.G. III,McClane, B.A. (2004).Fine mapping of the N-terminal cytotoxicity region of Clostridiumperfringens enterotoxin by site-directed mutagenesis. InfectionandImmunity,72,6914-6923.
Stringer, M.F., Watson, G.N. , Gilbert, R.J. , Wallace, J.G., Hassall, J.E. ,Tanner, E.I. , and Webbe, P.P. (1985).Fecal carriage of Closridiumperfringens.TheJournalofHygiene,95,277-288.
Sunagawa,H.,Takeshi, K.,Kameyama,K.,and Ando,Y. (1987).Incidence of Clostridiumperfringens in human feces and enterotoxin production and spor-germination of the isolates. JournaloftheFoodHygienicSocietyofJapan,28,119-124.
Tansuphasiri, U. (2001).Development of duplex PCR assay for rapid detection of enterotoxigenic isolates of Clostridiumperfringens. TheSoutheastAsianJournalofTropicalMedicineandPublicHealth, 32,105-113.
Van Damme-Jongsten, M. , Warnars, K. ,and Notermans, N. (1989).Cloning and sequencing of the Clostridiumperfringens enterotoxin gene. AntonievanLeevenhoek,56,181-190.
Varela, P.,Pollevick, G.D., Rivas, M.,Chinen, I., Binsztein, N.,Frasch, A.C.C. , and Ugalde, R.A. (1994).Direct detection of vibriocholera in stool samples. JournalofClinicalMicrobiology,32,1246-1248.
Widjojoatmodjo, M.N. ,Fluit, A.C. ,Torensma, R. ,Verdonk, G.P.H.T. ,andVerhoef,J. (1992).The magnetic immune polymerase chain reaction assay for direct detection ofSalmonella in fecal samples. JournalofClinicalMicrobiology,30,3195-3199.
Yamagishi, T., Serikawa, T.,Morita, R.,Nakamura, S., and Nishida, S. (1976).Persistent high numbers of Clostridiumperfringens in the intestines of Japanese aged adults. Japanese Journal of Microbioolgy, 20,397-403.
Asha, N.J., and Wilcox,M.H. (2002).Laboratory diagnosis of Clostridiumperfringens antibiotic-associated diarrhea. Journal ofMedicalMicrobiology, 51,891-894.
Berry, P.R.,Rodhouse, J.C.,Hughes, S.,Batholomew,B.A,and Gilbert, R.J. (1988).Evaluation of ELISA,RPLA and Vero cell assay for detecting Clostridiumperfringens enterotoxin in feacal specimens. JournalofClinicalPathologyl,41,458-461.
Eisgrüber, H., and Hauner, G. (2001).Minead beef heart associated with a Clostridiumperfringens food poisoning in a Munich old people’s home. ArchLebensmihelhyg, 52,63-66.
Fach,P.,andPopoff,M.R.(1997).Detection of enerotoxigenicClostridiumperfringens in food and fecal samples with a duplex PCR and the slide latex agglutination test. Applied and EnvironmentalMicrobiology, 63,4232-4236.
Gumerlock, P.H., Tang, Y.J., Weiss,J.B.,andSliva,J.Jr. (1993).Specific detection of toxigenic strains of Clostridiumdefficilein stool specimens. JournalofClinicalMicrobiology, 31,507-511.
Jay, J.M. (2000).Modern Food Microbiology(6thed.).Gaithersburg:AspenPuplishers.
Lahti, P., Heikinheimo, A., Johansson,T.,andKorkeala, H. (2008).Clostridiumperfringens type A strains carrying a plasmid-borne enterotoxin gene (genotype IS1151-cpe or IS1470-like-cpe) as a common cause of food poisoning .JournalofClinicalMicrobiology, 46,371-373.
Le Marc, Y., Plowman, J., Aldus, C.F., Munoz-Cuevas, M., Baranyi,J.,and Peck, M.W. (2008).Modelling the growth of Clostridiumperfringens during the cooling of bulk meat.InternationalJournalofFoodMicrobiology ,128,41-50.
Lukinmaa, S., Takkunen,E.,andSiitonen, A. (2002). Molecular epidemiology of Clostridiumperfringens related to food-borne outbreaks of disease in Finland from 1984 to 1999.AppliedandEnvironmentalMicrobiology,68,3744-3749.
Matches, J.R.,Liston,T.,andCurran, D. (1974).Clostridiumperfringens in the environment. AppliedMicrobiology, 28,655-660.
McClane, B.A. (2005).Clostridialenterotoxins. In: P .Dure(Eds),Handbook on Clostridia (pp. 385-406). BocaRaton:CRC Press.
McClane, B.A., Uzal,F.A., Miyakawa,M.F.,Lyerly,D.,and Wilkins, T. (2006).The enterotoxic Clostridia. In:M .Dworkin,S.Falkow,E. Rosen-burg,K.H.Schlcifer&E .Stackebrandt(Eds),The Prokaryotes,vol4(pp 698-752). New York:Springer-Verlag.
McClane, B.A. (2001).Clostridiumperfringens. In: M.P. Doyle,L.R.Beuchat&T.J. Montville (Eds),Food Microbiology:Fundamentals and Frontiers,2nded(pp351-37). ASM Press,Washington,D. C.
McClane,B.A.(2007) Clostridiumperfringens. In : M.P. Doyle & L.R. Beuchat (Eds),Food Microbiology: Fundamentals and Frontiers,3rded (pp433-444). ASM Press,Washington,D. C..
McClane,B.A.,andStrouse, R.J.(1984).Rapid detection of Clostridiumperfringens type A enterotoxin by enzyme-linked immunosorbent assay. JournalofClinicalMicrobiology, 19,112-115.
Miwa, N., Nishina, T. , Kubo,S. ,and Fujikura, K. (1996).Nested polymerase chain reaction for detection oflow levels of enterotoxigenicClosridiumperfringens in animal feces and meat . Journal ofVeterinaryMedical Science,58,197-203.
Monma, C.,Yanagawa, Y., Kusunoki, J., Kai, A.,Shingaki, M., Obata, H., Itoh, T., Ohta, K., and Kudoh, Y. (1994). Serotyping and enterotoxin production of heat-resistant spore-forming Clostridiumperfringens isolated from human feces.Annual Report ofTokyoMetropolitanResearchLaboratory ofPuplicHealth,45,11-15.
Nakamura,H., Ogasawara,J. , Monna, C., Suzuki,A.H., Kai,A.,Haruki,K.,andNishikawa, Y. (2003).Usefulness of a combination of pulsed-field gel electrophoresis and enrichment culture in laboratory invetigation of foodborne outbreak due to Clostridiumperfringens. DiagnosticMicrobiology andInfectiousDisease,47,471-475.
Petit, L.,Gilbert,M.,andPopoff, M.R. (1999) .Clostridiumperfringens: toxinotype and genotype. Trends in Microbiology, 7,104-110.
Rood, J.I. (1998).Virulance genes of Clostridiumperfringens. AnnualReviewofMicrobiology,52,333-360.
Smedley, J.G. III,McClane, B.A. (2004).Fine mapping of the N-terminal cytotoxicity region of Clostridiumperfringens enterotoxin by site-directed mutagenesis. InfectionandImmunity,72,6914-6923.
Stringer, M.F., Watson, G.N. , Gilbert, R.J. , Wallace, J.G., Hassall, J.E. ,Tanner, E.I. , and Webbe, P.P. (1985).Fecal carriage of Closridiumperfringens.TheJournalofHygiene,95,277-288.
Sunagawa,H.,Takeshi, K.,Kameyama,K.,and Ando,Y. (1987).Incidence of Clostridiumperfringens in human feces and enterotoxin production and spor-germination of the isolates. JournaloftheFoodHygienicSocietyofJapan,28,119-124.
Tansuphasiri, U. (2001).Development of duplex PCR assay for rapid detection of enterotoxigenic isolates of Clostridiumperfringens. TheSoutheastAsianJournalofTropicalMedicineandPublicHealth, 32,105-113.
Van Damme-Jongsten, M. , Warnars, K. ,and Notermans, N. (1989).Cloning and sequencing of the Clostridiumperfringens enterotoxin gene. AntonievanLeevenhoek,56,181-190.
Varela, P.,Pollevick, G.D., Rivas, M.,Chinen, I., Binsztein, N.,Frasch, A.C.C. , and Ugalde, R.A. (1994).Direct detection of vibriocholera in stool samples. JournalofClinicalMicrobiology,32,1246-1248.
Widjojoatmodjo, M.N. ,Fluit, A.C. ,Torensma, R. ,Verdonk, G.P.H.T. ,andVerhoef,J. (1992).The magnetic immune polymerase chain reaction assay for direct detection ofSalmonella in fecal samples. JournalofClinicalMicrobiology,30,3195-3199.
Yamagishi, T., Serikawa, T.,Morita, R.,Nakamura, S., and Nishida, S. (1976).Persistent high numbers of Clostridiumperfringens in the intestines of Japanese aged adults. Japanese Journal of Microbioolgy, 20,397-403.
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2013-09-30
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AL-MOLA, A. H., & AL-RAWI, A. M. (2013). Detection of Cpegene in Stool Samples of Food Poisoning Patients in Mosul/Iraq. Science Journal of University of Zakho, 1(2), 414–419. Retrieved from https://sjuoz.uoz.edu.krd/index.php/sjuoz/article/view/143
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