Rapid and sensitive detection of Enterobacter sakazakii by cross-priming amplification combined with immuno-blotting analysis

Zhao, Yuqing; Zhang, Xia; Zhang, Hongwei; Liu, Wei; Zheng, Wei; Huang, Xi‐Tai

doi:10.1016/j.mcp.2010.09.001

Cited by 38 publications

(20 citation statements)

References 24 publications

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“…This reaction was conducted at 63°C for 60 min and then stored at 4°C (Yulong et al, 2010). Six microliters final product of each CPA reaction was directly deposited onto each individual BESt strip (Ustar Biotech Co., Hangzhou, China) when CPA amplification was finished.…”

Section: Cpa Primers and Reaction Conditionsmentioning

confidence: 99%

“…Specifically, two testing primers in CPA are labeled with fluorescein isothiocyanate (FITC, a derivative of fluorescein) and biotin, respectively. The end of the amplicon labeled with biotin can bind to the colloidal gold, while the other end labeled with FITC can hybridize with the anti-FITC antibody located on the strip test line (Yulong et al, 2010). This subsequently results in a red color of colloidal gold accumulating at the test line for direct visualization within 10 min.…”

Section: Introductionmentioning

confidence: 99%

“…In addition, CPA coupled with immunoblotting analysis was successfully applied to detect Cronobacter sakazakii (Yulong et al, 2010). The objective of the current study was to develop a new CPA assay combined with immunoblotting analysis for the rapid detection of Y. enterocolitica in milk powders.…”

Section: Introductionmentioning

confidence: 99%

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Detection of Yersinia enterocolitica in milk powders by cross-priming amplification combined with immunoblotting analysis

Zhang

Feng

Zhao

et al. 2015

International Journal of Food Microbiology

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Section: Cpa Primers and Reaction Conditionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Detection of Yersinia enterocolitica in milk powders by cross-priming amplification combined with immunoblotting analysis

Zhang

Feng

Zhao

et al. 2015

International Journal of Food Microbiology

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“…A vertical flow (VF) hybridization-based nucleic acid detection strip cassette was designed to visually detect double-labeled amplicons in 5 to 10 min (8). CPA coupled with a VF visualization strip has been developed successfully to detect specific DNAs from several bacterial pathogens, including Mycobacterium tuberculosis and Enterobacter sakazakii (3,17). In this study, CPA was modified to include reverse transcription-CPA (RT-CPA) technology to rapidly detect SFTSV-specific RNA.…”

mentioning

confidence: 99%

Detection of Severe Fever with Thrombocytopenia Syndrome Virus by Reverse Transcription–Cross-Priming Amplification Coupled with Vertical Flow Visualization

Cui

et al. 2012

J Clin Microbiol

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A virus known as severe fever with thrombocytopenia syndrome virus (SFTSV) was recently identified as the etiological agent of severe fever with thrombocytopenia syndrome (SFTS) in China. Reliable laboratory detection and identification of this virus are likely to become clinically and epidemiologically desirable. We developed a nearly instrument-free, simple molecular method which incorporates reverse transcription-cross-priming amplification (RT-CPA) coupled with a vertical flow (VF) visualization strip for rapid detection of SFTSV. The RT-CPA-VF assay targets a conserved region of the M segment of the SFTSV genome and has a limit of detection of 100 copies per reaction, with no cross-reaction with other vector-borne bunyaviruses and bacterial pathogens. The performance of the RT-CPA-VF assay was determined with 175 human plasma specimens collected from 89 clinically suspected SFTS patients and 86 healthy donors. The sensitivity and specificity of the assay were 94.1% and 100.0%, respectively, compared with a combination of virus culture and real-time RT-PCR. The entire procedure, from specimen processing to result reporting, can be completed within 2 h. The simplicity and nearly instrument-free platform of the RT-CPA-VF assay make it practical for point-of-care testing.

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“…An ideal method for current molecular biology diagnosis should be simple and feasible to use in any laboratory conditions. Therefore, loop-mediated amplification (LAMP) together with other isothermal techniques including cross-priming amplification or helicase-dependent amplification are exceptionally interesting and useful (Vincent et al 2004;Jeong et al, 2009;Yulong et al, 2010;Xu et al, 2012;Yang et al, 2014;Vincent et al, 2004). The previous report of LAMP application in detection of viral agents of waterfowl showed its robustness in detection of Derzsy's disease virus (GPV; Yang et al, 2010), MDPV (Ji et al, 2010), GCV (Woźniakowski et al, 2012) or DEV (Ji et al, 2009).…”

Section: Discussionmentioning

confidence: 99%

Visual detection of goose haemorrhagic polyomavirus in geese and ducks by loop-mediated isothermal amplification

Woźniakowski

Tarasiuk

2015

Avian Pathology

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Goose haemorrhagic polyomavirus (GHPV) is an aetiological agent of haemorrhagic nephritis and enteritis of geese occurring in geese (Anser anser). GHPV may also infect Muscovy ducks (Carina mochata) and mule ducks. Early detection of GHPV is important to isolate the infected birds from the rest of the flock thus limiting infection transmission. The current diagnosis of haemorrhagic nephritis and enteritis of geese is based on virus isolation, histopathological examination, haemagglutination inhibition assay, ELISA and polymerase chain reaction (PCR). Recently, real-time PCR assay was developed which considerably improved detection of GHPV. In spite of many advantages, these methods are still time-consuming and inaccessible for laboratories with limited access to ELISA plate readers or PCR thermocyclers. The aim of our study was to develop loop-mediated isothermal amplification (LAMP) that may be conducted in a water bath. Two pairs of specific primers complementary to VP1 gene of GHPV were designed. The results of GHPV LAMP were recorded under ultraviolet light. Our study showed LAMP was able to specifically amplify VP1 fragment of a GHPV without cross-reactivity with other pathogens of geese and ducks. LAMP detected as little as 1.5 pg of DNA extracted from a GHPV standard strain (150 pg/µl). The optimized LAMP was used to examine 18 field specimens collected from dead and clinically diseased geese and ducks aged from 1 to 12 weeks. The positive signal for GHPV was detected in three out of 18 (16.6%) specimens. These results were reproducible and consistent with those of four real-time PCR. To the best of our knowledge this is the first report on LAMP application for the GHPV detection.

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Rapid and sensitive detection of Enterobacter sakazakii by cross-priming amplification combined with immuno-blotting analysis

Cited by 38 publications

References 24 publications

Detection of Yersinia enterocolitica in milk powders by cross-priming amplification combined with immunoblotting analysis

Detection of Yersinia enterocolitica in milk powders by cross-priming amplification combined with immunoblotting analysis

Detection of Severe Fever with Thrombocytopenia Syndrome Virus by Reverse Transcription–Cross-Priming Amplification Coupled with Vertical Flow Visualization

Visual detection of goose haemorrhagic polyomavirus in geese and ducks by loop-mediated isothermal amplification

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