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Rapid and quantitative detection of Shiga toxin1 and Shiga toxin2 based on multiple targets UPT-LF assay
Wei, Qiaozhen1,2; Hu, Qiushi1; Shi, Fengjuan3; Li, Shuang1; Sun, Chongsi1; Zhang, Huicong1; Xue, Lei1; Feng, QiuXia1; Dong, Jinying1; Jiao, Yongjun3; Zhou, Lei1
2020-07-19
Source PublicationENGINEERING IN LIFE SCIENCES
ISSN1618-0240
Pages10
AbstractShiga toxin-producingEscherichia coli(STEC) infection causes a series of diseases that are highly pathogenic and deadly in humans and animals, seriously endangering public health. Of the pathogenic factors within STEC, the two groups ofShiga toxin(Stx) consisting Stx1 and Stx2 plays a prominent role in the pathogenesis of STEC infection. In this study, we developed single-target up-converting phosphor technology-based lateral flow assay (Stx-UPT-LFA) for the rapid detection of Stx1 and Stx2, respectively, and also developed a dual-target Stx1/2-UPT-LFA based on single-target strips to detect of Stx1 and Stx2 at the meantime within 20 min. We choose the purified Stx1 and Stx2 standard samples, and the optimum monoclonal antibody (namely 8E7-E6, 2F6-F8 for Stx1 and S1D8, S2C4 for Stx2) were selected for use in Stx-UPT-LFA in double-antibody-sandwich mode. The sensitivities of single-target Stx-UPT-LFA for both Stx1 and Stx2 were 1 ng mL(-1)with accurate quantitation ranges of 1-1000 ng mL(-1)and 1-800 ng mL(-1)respectively. No false-negative result was found in the Stx2-UPT-LFA even with a high-test concentration up to 1000 ng mL(-1). Meanwhile, both targets detection sensitivities for dual-target Stx1/2-UPT-LFA were 5 ng mL(-1), and accurate quantitation ranges were 5-1000 ng mL(-1)and 5-800 ng mL(-1)for standard Stx1 and Stx2 solutions without cross-interference between two targets. Both techniques showed good linearities, with a linear fitting coefficient of determination(r) of 0.9058-0.9918. Therefore, the UPT-LFA could realize simultaneous detection for multiple targets on a single strip and thus to quickly determine the type of infectious Stxs. In addition, the single-target Stx1-UPT-LFA and Stx2-UPT-LFA showed excellent specificity to six toxins, even at high concentrations of 1000 ng mL(-1). In conclusion, the developed Stx-UPT-LFA allows the rapid, quantitative, reliable and simultaneous detection of Stx1 and Stx2 within 20 min, providing an alternative method for clinical diagnosis of STEC infection.
Keyworddual-target lateral flow assay Shiga toxin single-target up-converting phosphor technology
DOI10.1002/elsc.202000031
Language英语
WOS KeywordESCHERICHIA-COLI INFECTIONS ; IMMUNOASSAY ; DIAGNOSIS ; PATHOGENS ; BIOMARKER ; DISEASE ; LABELS
Funding ProjectNational Key Research and Development Program[2019YFC1606600] ; National Key Research and Development Program[2019YFC1606602] ; Innovation Program of National Defense Science and Technology[19-H863-18-ZD-011-004-03] ; Beijing Nova Programme Interdisciplinary Cooperation Project[Z191100001119005]
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
Funding OrganizationNational Key Research and Development Program ; Innovation Program of National Defense Science and Technology ; Beijing Nova Programme Interdisciplinary Cooperation Project
WOS IDWOS:000549859200001
PublisherWILEY
Citation statistics
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/41414
Collection中国科学院过程工程研究所
Corresponding AuthorJiao, Yongjun; Zhou, Lei
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, PLA Key Lab Biopharmaceut Prod & Formulat Engn, Beijing 100190, Peoples R China
2.Anhui Med Univ, Dept Blood Transfus, Affiliated Hosp 2, Hefei, Peoples R China
3.Jiangsu Prov Ctr Dis Prevent & Control, Inst Pathogen Microbiol, Nanjing 210009, Peoples R China
Recommended Citation
GB/T 7714
Wei, Qiaozhen,Hu, Qiushi,Shi, Fengjuan,et al. Rapid and quantitative detection of Shiga toxin1 and Shiga toxin2 based on multiple targets UPT-LF assay[J]. ENGINEERING IN LIFE SCIENCES,2020:10.
APA Wei, Qiaozhen.,Hu, Qiushi.,Shi, Fengjuan.,Li, Shuang.,Sun, Chongsi.,...&Zhou, Lei.(2020).Rapid and quantitative detection of Shiga toxin1 and Shiga toxin2 based on multiple targets UPT-LF assay.ENGINEERING IN LIFE SCIENCES,10.
MLA Wei, Qiaozhen,et al."Rapid and quantitative detection of Shiga toxin1 and Shiga toxin2 based on multiple targets UPT-LF assay".ENGINEERING IN LIFE SCIENCES (2020):10.
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