Abstract:
:The specific identification of verocytotoxin (VT)-producing Escherichia coli (VTEC) requires the detection of VTs in bacterial culture filtrates or the detection of genes encoding these toxins in bacterial cells by specific DNA probes. The standard method for detecting these toxins involves a time-consuming, labor-intensive, and expensive cytotoxicity assay. We have developed a specific, highly sensitive receptor-specified enzyme-linked immunosorbent assay (RELISA) to detect VT1, one of at least two VTs implicated in human disease. The assay is based on the affinity of VT1 for the glycolipid globotriosyl ceramide (Gb3). Gb3 was de-N-acylated to yield lyso-Gb3, which is more polar but retains VT1 binding. Lyso-Gb3 was used to sensitize microdilution plates to bind VT1 for subsequent immunodetection. This RELISA was used to detect VT1 in the culture supernatant of a variety of bacteria of known VT status. The assay was compared with the highly sensitive cell cytotoxicity assay for their abilities to detect VT. The RELISA was as sensitive as the cytotoxicity assay and, in a blind study, 100% specific. This assay will provide a quick, specific, efficient adjunct to the diagnosis and epidemiological study of VTEC infections and their relationship to human disease.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Basta M,Karmali M,Lingwood Cdoi
10.1128/JCM.27.7.1617-1622.1989subject
Has Abstractpub_date
1989-07-01 00:00:00pages
1617-22issue
7eissn
0095-1137issn
1098-660Xjournal_volume
27pub_type
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
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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:10.1128/JCM.16.1.53-59.1982
更新日期:1982-07-01 00:00:00
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journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.33.1.126-130.1995
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abstract::A real-time quantitative PCR assay has been developed to measure human herpesvirus 6 (HHV-6) DNA in biological specimens. The assay sensitivity was 10 copies of DNA per well, with a linear dynamic range of 10 to 10(7) copies of HHV-6 DNA. Intra- and interassay variations were, respectively, 0.88 and 0.8% for samples c...
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doi:10.1128/jcm.39.9.3197-3199.2001
更新日期:2001-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.3.484-489.1993
更新日期:1993-03-01 00:00:00
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doi:10.1128/JCM.35.10.2665-2669.1997
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.12.2659-2660.1988
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journal_title:Journal of clinical microbiology
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pub_type: 杂志文章
doi:10.1128/JCM.33.8.2016-2021.1995
更新日期:1995-08-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 临床试验,杂志文章
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更新日期:2005-01-01 00:00:00
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