MULTIPLEX PCR ASSAY FOR TOXINOTYPING CLOSTRIDIUM PERFRINGENS ISOLATES OBTAINED FROM HERBS

Authors

  • Piyada Wangroongsarb National Institute of Health, Department of Medical Sciences, Ministry of Public Health.
  • Chutima Jittaprasatsin 1National Institute of Health, Department of Medical Sciences, Ministry of Public Health.
  • Thanitchai Kamthalang National Institute of Health, Department of Medical Sciences, Ministry of Public Health.
  • Nattapong Cheunban National Institute of Health, Department of Medical Sciences, Ministry of Public Health.
  • Somchai Sangkitporn National Institute of Health, Department of Medical Sciences, Ministry of Public Health.
  • Thayat Sriyapai Major in Environment, Faculty of Environmental Culture and Ecotourism, Srinakharinwirot University.

Keywords:

Clostridium perfringens, Multiplex PCR, Enterotoxin, Herbs

Abstract

Two hundred and forty samples of herbs were collected by the Department of Medical Sciences from 2012-2019 to detect Clostridium perfringens by conventional culture method and molecular typing by multiplex PCR. Four major toxin genes (cpa, cpb, etx and iA) and an enterotoxin gene (cpe) from C. perfringens isolates were detected. In this study, C. perfringens isolates were detected in 112 (46.67 %) herb samples by culture method, all of which were genotyped as C. perfringens type A. However, 6 of all type A isolates were positive for the enterotoxin-encoding gene (cpe gene). Enterotoxin-producing C. perfringens were found in 4 types of herbs including wasting disease drug (2 samples), veld grape (1 sample), sweet leaf (2 samples) and turmeric (1 sample). The monitoring of C. perfringens strains by molecular techniques for typing and identifying toxin-producing isolates is useful for the epidemiology data and control the quality of herbal products to meet the standards legal requirements.

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References

Van Immerseel, F., De Buck, J., Pasmans, F., Huyghebaert, G., Haesebrouck, F., Ducatelle, R. (2004). Clostridium perfringens in poultry: an emerging threat for animal and public health. Avian Pathology, 33(6), 537-549.

Petit, L., Gibert, M., Pofoff, M.R. (1999). Clostridium perfringens: toxinotype and genotype. Trends Microbiology, 7, 104-110.

. Gokce, H.I., Genc, O., Sozmen, M., Gokce, G. (2007). Determination of Clostridium perfringens toxin-types in sheep with suspected enterotoxemia in Kars Province, Turkey. Turkish Journal of Veterinary and Animal Sciences, 31(5), 355-360.

Songer, J.G, Meer R.R. (1996). Genotyping of Clostridium perfringens by polymerase chain reaction is a useful adjunct to diagnosis of clostridial enteric disease in animals. Anaerobe, 2, 197-203.

Garmory, H.S, Chanter, N., French, N.P., Bueschel, D., Songer, J.G., Titball, R. W. (2000). Occurrence of Clostridium perfringens beta2-toxin amongst animals, determined using genotyping and subtyping PCR assays. Epidemiology and Infection, 124, 61-67.

Miki, Y., Miyamoto, K., Kaneko-Hirano, I., Fujiuchi, K., Akimoto S. (2008). Prevalence and characterization of enterotoxin gene-carrying Clostridium perfringens isolates from retail meat products in Japan. Applied and Environmental Microbiology, 74, 5366-5372.

Al Khaldi, S.F, Myers, K.M., Rasooly, A., Chizhikov, V. (2004). Genotyping of Clostridium perfringens toxins using multiple oligonucleotide microarray hybridization. Molecular and Cellular Probes, 18, 359-367.

Albini, S., Brodard, I., Jaussi, A., Wollschlaeger, N., Frey, J., Miserez, R., Abril, C. (2008). Real-time multiplex PCR assays for reliable detection of Clostridium perfringens toxin genes in animal isolates. Veterinary Microbiology, 127, 179-185.

Department of Medical Sciences. (2009). Thai herbal pharmacopoeia Vol. III. Nonthaburi: Department of Medical Sciences, Ministry of Public Health.

Chaiyawat, C., Jamtaweekul, J., Wongpentak, S., Prachyaporn, I. (2014). Safety of herbal medicines in the National List. Bulletin of the Department of Medical Sciences, 56(3), 123-134.

Dowell, V.R. Jr, Hawkins, T.M. (1987). Laboratory methods in anaerobic bacteriology CDC Laboratory Manual. Atlanta: DHEW Publication, p. 8272.

Holdeman, L.V., Cato, E.P., & Moore, W.E.C. (1977). Anaerobe Laboratory Manual. 4th ed. Blacksburg: Virginia Polytechnic Institute and State University. p. 156.

Sambrook, J., Fritsch, E.F., Maniatis, T. (1989). Molecular cloning: a laboratory manual. 2nd ed. Cold Spring Harbor: Cold Spring Harbor Laboratory Press. p. 1989

Heikinheimo, A., Korkeala, H. (2005). Multiplex PCR assay for toxinotyping Clostridium perfringens isolates obtained from Finnish broiler chickens. Letters in Applied Microbiology, 40(6), 407-411.

Meer, R.R., Songer, J.G. (1997). Multiplex polymerase chain reaction assay for genotyping Clostridium perfringens. American Journal of Veterinary Research, 58(7), 702-705.

Bureau of Drug and Narcotics, Department of Medical Sciences, Ministry of Public Health. (2011). Annual report in 2011. Nonthaburi: Ministry of Public Health.

Jepson, M., Titball, R.W. (2000). Structure and function of clostridial phospholipase C. Microbes and Infection, 2, 1277-1284.

Krakauer, T., Fleischer, B., Stevens, D.L., McClane, B.A., Stiles, B.G. (1997). Clostridium perfringens enterotoxin lacks superantigenic activity but induces an interleukin-6 response from human peripheral blood mononuclear cells. Infection and Immunity, 65(8), 3485-3488.

McClane, B.A. (1996). An overview of Clostridium perfringens enterotoxin. Toxicon, 34(11-12), 1335-1343.

Engström, B.E., Fermér, C., Lindberg, A., Saarinen, E., Båverud, V., Gunnarsson, A. (2003). Molecular typing of isolates of Clostridium perfringens from healthy and diseased poultry. Veterinary Microbiology, 94, 225-235.

Deguchi, A., Miyamoto, K., Kuwahara, T., Miki, Y., Kaneko, I., & Li, J., McClane B.A., Akimoto, S. (2009). Genetic characterization of type A enterotoxigenic Clostridium perfringens strains. PLoS ONE. 4(5), e5598.

Published

2021-12-30