Characterization of Giardia duodenalis by polymerase-chain-reaction fingerprinting (original) (raw)

Abstract

The development of a polymerase chain reaction (PCR) based fingerprinting method for the characterization of Giardia duodenalis isolates is described. The method uses three different PCRs; one is specific for the A (“Polish”) major group of G. duodenalis isolates, another is specific for the B (“Belgian”) group of isolates; and one amplifies a fragment of the glutamate dehydrogenase gene present in all G. duodenalis isolates. The PCRs perform highly sensitively on DNA from cultured trophozoites. Isolates were further characterized by restriction-fragment-length polymorphism (RFLP) analysis of the PCR products. In this way, representative isolates from the A and B groups could be grouped together into a number of subgroups. The stability of the genotypes with time and the reproducibility of the two methods were tested on cloned and subcloned lines from a number of isolates and proved to be highly satisfactory. The PCR/RFLP method was evaluated on cysts derived from a number of human patients. It is concluded that the PCR fingerprinting method described in this paper provides a reliable characterization method for Giardia isolates and has the potential to be used as a direct method of typing G. duodenalis cysts from feces.

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Authors and Affiliations

  1. Microbiological Laboratory of Health Protection, National Institute of Public Health and the Environment, P.O. Box 1, 3720 BA Bilthoven, The Netherlands e-mail: wieger.homan@rivm.nl, Tel.: +31 30 2742909, Fax: +31 30 2744434, , , , , , NL
    Wieger L. Homan, Margriet Gilsing, Hafida Bentala & Louis Limper
  2. Department of the Science of Food of Animal Origin, University of Utrecht, Utrecht, The Netherlands, , , , , , NL
    Frans van Knapen

Authors

  1. Wieger L. Homan
  2. Margriet Gilsing
  3. Hafida Bentala
  4. Louis Limper
  5. Frans van Knapen

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Received: 7 January 1998 / Accepted: 7 April 1998

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Homan, W., Gilsing, M., Bentala, H. et al. Characterization of Giardia duodenalis by polymerase-chain-reaction fingerprinting.Parasitol Res 84, 707–714 (1998). https://doi.org/10.1007/s004360050474

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