Detection of Viable Cryptosporidium parvum by Real-Time RT-PCR |
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학술지명 한국환경분석학회
저자 이규철,이상태,이정임,남세희
발표일 2007-11-14
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Cryptosporidium parvum (C. parvum) is one of the waterborne protozoan parasites that are well known to cause Cryptosporidiosis. These parasites exist in the water system in very low densities and are resistant to many environmental stresses, thereby necessitating the development of a very sensitive detection method to detect them. In this study, we developed a real-time reverse transcription (RT)-polymerase chain reaction (PCR) method to detect the mRNA of the hsp70 gene for the detection of viable C. parvum. In the results, it was shown that the detection sensitivity for hsp70 was equal to that of the RT-PCR for hsp70 which had been developed. Also, it was possible to observe a 10-fold increase of the detection sensitivity when hsp70 mRNA was induced by giving heat shock, compared with that of the case that was not subjected to heat shock. In conclusion, the real-time PCR method for the hsp70 gene of C. parvum developed in this study is as sensitive as the RT-PCR method, as well as it makes quantification possible. Also, by using primers and probes for hsp70 gene, it was possible to detect viable C. parvum as well as achieve about 10-fold increase in detection sensitivity. Further studies are needed to detect viable Giardia lamblia, known to be an important waterborne parasite, by using real-time RT-PCR. |