Abstract：
:The performance of polymerase chain reaction (PCR) procedures for the detection of Cryptosporidium parvum HNJ-1 strain (genotype II) oocysts purified from mice using published protocols was evaluated. Oocysts were concentrated from fecal samples of infected severe combined immunodeficiency (SCID) mice by sucrose flotation and were then purified by immunomagnetic separation method. The genotype of C. parvum was established as type II by restriction fragment length polymorphism (RFLP) analysis. Water samples were spiked with different numbers of oocysts, determined by limiting dilution. Genomic DNA was extracted and used for PCR assays targeting various Cryptosporidium species genes (Beta-Tubulin, COWP, 70 kDa HSP, SSU rRNA, ITS1, TRAP-C1 and TRAP-C2 gene). DNA from oocyst numbers of more than 1 x 10(4) was detected using each of the primers. However, when using lower oocyst numbers, the tools based on 9 of the 16 different primer assays gave sufficient results. Assays using the remaining seven primers gave less than satisfactory results. A new primer set, named VKSS-F1/2 and VKSS-R1/2, that target the 18 SSU rRNA gene of C. parvum was constructed and applied. The VKSS-F1/2 and VKSS-R1/2 assays amplified DNA isolated from spiked samples in 206 of 211 trials (97.6%). This illustrates the difficulty of detecting low numbers of Cryptosporidium spp. oocysts by molecular methods when working with environmental samples.
journal_name
Parasitol Resjournal_title
Parasitology researchauthors
Leetz AS,Sotiriadou I,Ongerth J,Karanis Pdoi
10.1007/s00436-007-0567-ysubject
Has Abstractpub_date
2007-09-01 00:00:00pages
951-62issue
4eissn
0932-0113issn
1432-1955journal_volume
101pub_type
杂志文章abstract：:To date classification and differentiation of Alaria spp. are based largely on external characteristics and comparative morphology of adult flukes. The accurate differentiation between various Alaria spp. mesocercariae is indeed difficult because there are only few data on morphological and morphometrical features of ...
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journal_title：Parasitology research
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doi：10.1007/BF00931056
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journal_title：Parasitology research
pub_type： 杂志文章,评审
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更新日期：2016-05-01 00:00:00
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pub_type： 杂志文章
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更新日期：2008-03-01 00:00:00
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journal_title：Parasitology research
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更新日期：2012-02-01 00:00:00
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journal_title：Parasitology research
pub_type： 杂志文章
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更新日期：2007-08-01 00:00:00
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journal_title：Parasitology research
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doi：10.1007/s00436-007-0566-z
更新日期：2007-09-01 00:00:00
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journal_title：Parasitology research
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