Sensitive detection of Cryptosporidium oocysts in environmental water samples by reverse transcription-PCR.
نویسندگان
چکیده
Pupply. With a reverse transcription-PCR (RT-PCR) amplifylng heat shock protein 70 (hsp70) gene, we were able to detect two differenttypes of viable Cyyptosporidium oocysts from the environment. Water specimens were filtered under positive pressure through a 3-Elm polycarbonate filter (Nuclepore filter, Whatman, Maidstone, England). The trapped materials were detached from the filter by sonication, and concentrated by
منابع مشابه
Sensitive and rapid detection of viable Giardia cysts and Cryptosporidium parvum oocysts in large-volume water samples with wound fiberglass cartridge filters and reverse transcription-PCR.
We recently described a reverse transcription-PCR (RT-PCR) for detecting low numbers of viable Cryptosporidium parvum oocysts spiked into clarified environmental water concentrates. We have now modified the assay for direct analysis of primary sample concentrates with simultaneous detection of viable C. parvum oocysts, Giardia cysts, and a novel type of internal positive control (IPC). The IPC ...
متن کاملDetection and enumeration of Cryptosporidium oocysts in environmental water samples by Real-time PCR assay
Introduction: The protozoan parasite, Cryptosporidium Spp., widely spreads in both raw and drinking waters. It is the causative agents of waterborne diarrhea and gastroenteritis in the world. In the present study, a molecular assay was used for the detection and quantification of Cryptosporidium oocysts in environmental water samples. Materials and methods: Thirty surface water samples wer...
متن کاملDetection of viable Cryptosporidium parvum in soil by reverse transcription-real-time PCR targeting hsp70 mRNA.
Extraction of high-quality mRNA from Cryptosporidium parvum is a key step in PCR detection of viable oocysts in environmental samples. Current methods for monitoring oocysts are limited to water samples; therefore, the goal of this study was to develop a rapid and sensitive procedure for Cryptosporidium detection in soil samples. The efficiencies of five RNA extraction methods were compared (mR...
متن کاملDevelopment of a PCR protocol for sensitive detection of Cryptosporidium oocysts in water samples.
The development of a reliable method of using PCR for detection of Cryptosporidium oocysts in environmental samples with oligonucleotide primers which amplify a portion of the sequence encoding the small (18S) subunit of rRNA producing a 435-bp product was demonstrated. The PCR assay was found to provide highly genus-specific detection of Cryptosporidium spp. after release of nucleic acids from...
متن کاملEvaluation of immunomagnetic separation and the sucrose flotation methods coupled with immunofluorescence or PCR for detection of Cryptosporidium and Giardia (oo)cysts in water samples
Introduction: Detection of Cryptosporidium and Giardia parasites in water samples is usually performed by US Environmental Protection Agency 1623 method. Nevertheless, the USEPA1623 method still need improvement, to prevent and control the water borne parasitic disease. Therefore, we undertook the present study. Materials and methods: Totally 48 surface water samples were collected. Four sampl...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
- Japanese journal of infectious diseases
دوره 54 3 شماره
صفحات -
تاریخ انتشار 2001