Abstract:
:We report the development of a real-time PCR assay for the quantitative detection of Neospora caninum in infected host tissues. The assay uses the double-stranded DNA-binding dye SYBR Green I to continuously monitor product formation. Oligonucleotide primers were designed to amplify a 76-bp DNA fragment corresponding to the Nc5 sequence of N. caninum. A similar method was developed to quantify the 28S rRNA host gene in order to compare the parasite load of different samples and to correct for the presence of potential PCR-inhibiting compounds in the DNA samples. A linear quantitative detection range of 6 logs with a calculated detection limit of 10(-1) tachyzoite per assay was observed with excellent linearity (R(2) = 0.998). Assay specificity was confirmed by using DNA from the closely related parasite Toxoplasma gondii. The applicability of the technique was successfully tested in a variety of host brain tissues: (i) aborted bovine fetuses classified into negative or positive Neospora-infected animals according to the observation of compatible lesions by histopathological study and (ii) experimentally infected BALB/c mice, divided into three groups, inoculated animals with or without compatible lesions and negative controls. All samples were also tested by ITS1 Neospora nested PCR and a high degree of agreement was shown between both PCR techniques (kappa = 0.86). This technique represents a useful quantitative diagnostic tool to be used in the study of the pathogenicity, immunoprophylaxis, and treatment of Neospora infection.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Collantes-Fernández E,Zaballos A,Alvarez-García G,Ortega-Mora LMdoi
10.1128/jcm.40.4.1194-1198.2002subject
Has Abstractpub_date
2002-04-01 00:00:00pages
1194-8issue
4eissn
0095-1137issn
1098-660Xjournal_volume
40pub_type
杂志文章abstract::We describe two rapid, simple, and reliable procedures for routine purification of hepatitis B virus (HBV) DNA from serum. HBV DNA could be purified from 24 serum samples in 1.5 to 2 h and was recovered in the initial reaction vessel. Both procedures have in common that HBV DNA is complexed with silica particles in th...
journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 临床试验,杂志文章
doi:
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02036-09
更新日期:2010-04-01 00:00:00
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journal_title:Journal of clinical microbiology
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pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.32.9.2092-2098.1994
更新日期:1994-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.5.970-972.1986
更新日期:1986-05-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.9.2231-2235.1996
更新日期:1996-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.16.4.622-626.1982
更新日期:1982-10-01 00:00:00
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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pub_type: 杂志文章
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更新日期:2001-03-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:1984-08-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:2003-10-01 00:00:00
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pub_type: 杂志文章
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更新日期:2004-11-01 00:00:00
abstract::DNA fingerprint patterns of 156 Cryptococcus neoformans isolates (26 AIDS patients, 46 non-AIDS patients, and 40 environmental sources) from both varieties (126 C. neoformans var. neoformans and 30 C. neoformans var. gattii isolates) and from seven countries were analyzed by using the DNA probe UT-4p. Nine and twelve ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.7.1807-1814.1995
更新日期:1995-07-01 00:00:00
abstract::The Autobac IDX system (General Diagnostics, Warner-Lambert Co., Morris Plains, N.J.) for rapid, semiautomated identification of gram-negative bacilli was compared with the identification methods in routine use in four laboratories. The study included 1,515 organisms representing 30 species of enteric and nonenteric b...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.19.4.529-533.1984
更新日期:1984-04-01 00:00:00
abstract::We studied whether the Etest can be used as an alternative to agar dilution to determine antimicrobial susceptibilities of ceftriaxone, cefixime, and cefpodoxime in Neisseria gonorrhoeae surveillance. One hundred fifteen clinical and laboratory isolates of N. gonorrhoeae were tested following the Clinical Laboratory I...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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更新日期:2014-05-01 00:00:00