TITLE

DETECTING LIVE AND DEAD CELLS OF PECTOBACTERIUM ATROSEPTICUM BASED ON IMMUNOMAGNETICS SEPARATION AND STAINING

AUTHOR(S)
Sledz, W.; Adamowska, A.; Piosik, J.; Lojkowska, E.
PUB. DATE
November 2012
SOURCE
Journal of Plant Pathology;Nov2012, Vol. 94 Issue 3, p681
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
An assay that combines immunomagnetic separation (IMS) and staining by the LIVE/DEAD BacLightâ„¢ Bacterial Viability Kit was developed for the rapid detection and identification of living and dead cells of the plant-pathogenic bacterium Pectobacterium atrosepticum (Pba) in potato tuber tissue. In the development of this IMS/LD assay, bacterial suspensions in buffer or potato peel extract were spiked with living and dead cells of Pba and incubated with paramagnetic beads coated with Pba-specific antibodies. Beads with Pba cells were recovered from the suspensions with a magnet and the immobilized cells were stained with a mixture of Syto9 and propidium iodide. The collected cells were analysed by fluorescence microscopy. The assay required minimal sample preparation (e.g. no wash steps were required) and could be completed within 1 h. The detection limit was 104 and 104 cells per ml for M-280 and M-450 paramagnetic beads, respectively, which is similar to that previously reported for ELISA. The specificity and sensitivity of the IMS/LD assay were similar regardless of whether Pba cells were in potato peel extracts or in buffer. The assay also allowed the differentiation of live and dead Pba cells, which is important in predicting possible losses caused by Pba to potato crops.
ACCESSION #
86441973

 

Related Articles

  • Newly Established Monoclonal Antibody Diagnostic Assays for Schistosoma mansoni Direct Detection in Areas of Low Endemicity. Grenfell, Rafaella Fortini Queiroz; Coelho, Paulo Marcos Zech; Taboada, Diana; de Mattos, Ana Carolina Alves; Davis, Ruth; Harn, Donald A. // PLoS ONE;Jan2014, Vol. 9 Issue 1, p1 

    Background: Current available methods for diagnosis of schistosomiasis mansoni lack sufficient sensitivity, which results in underreporting of infectious in areas of low endemicity. Methodology/Principal Findings: We developed three novel diagnostic methodologies for the direct detection of...

  • Specific detection of foot-and-mouth disease serotype Asia 1 virus by carboxyl-magnetic beads conjugated with single-domain antibody. Shunli Yang; Shuanghui Yin; Youjun Shang; Di Wang; Weimin Ma; Jijun He; Jianhong Guo; Jianping Cai; Xiangtao Liu // BMC Biotechnology;9/15/2015, Vol. 15 Issue 1, p1 

    Background: Immunomagnetic nanobead (IMNB) labeled with specific antibody, has been demonstrated to be useful for the capturing and detection of viruses. Results: In this study, we developed an imunomagnetic bead based on carboxyl-magnetic beads (MNB) labeled with a single-domain antibody (sdAb)...

  • Lateral-flow enzyme immunoconcentration for rapid detection of Listeria monocytogenes. Cho, Il-Hoon; Irudayaraj, Joseph // Analytical & Bioanalytical Chemistry;Apr2013, Vol. 405 Issue 10, p3313 

    Lateral-flow enzyme immunochromatography coupled with an immunomagnetic step was developed for rapid detection of Listeria monocytogenes in food matrices. The target bacteria was first separated and concentrated by magnetic nanoparticles containing the enzyme and directly applied to the assay...

  • Development of Lectin-Linked Immunomagnetic Separation for the Detection of Hepatitis A Virus. Sang-Mu Ko; Kwon, Joseph; Vaidya, Bipin; Jong Soon Choi; Hee-Min Lee; Myung-Joo Oh; Hyeun-Jong Bae; Se-Young Cho; Kyung-Seo Oh; Duwoon Kim // Viruses (1999-4915);Mar2014, Vol. 6 Issue 3, p1037 

    The accuracy and sensitivity of PCR-based methods for detection of hepatitis A virus (HAV) are dependent on the methods used to separate and concentrate the HAV from the infected cells. The pH and ionic strength affect the binding affinity of the virus to cells. In this study, we initially...

  • Application review of immunology technology in detection for foodborne microorganism. HU Jin-qiang; LEI Jim-ting; ZHAN Li-juan; ZONG Wei; BAI Yan-hong; JING Jian-zhou; SUN Xin-cheng; DONG Cai-wen // Journal of Zhengzhou University of Light Industry, Natural Scien;Jun2014, Vol. 29 Issue 3, p7 

    Applification of enzyme-linked immunosorbent assay (ELISA) immunochromatography (IC), immunomagnetic separation (IMS), enzyme-linked fluorescent immunoassay (ELFIA), chemiluminescence immunoassay (CLIA) as well as immunosensor (IS) in the detection of food safety has been summarized. Moreover,...

  • Serology: Now and in the Future. Diiulio, Renee // Clinical Lab Products;Sep2010, Vol. 40 Issue 9, p30 

    This article explores trends in the field of serology testing. It identifies three main methodologies in use for serology testing: indirect immunofluorescent microscopy (IFA), enzyme-linked immunoabsorbent assays (ELISAs) and the multiplex technology. It also stresses the need for greater...

  • Comparison of Different Methods of Detection of Enteric Pathogenic Protozoa. Ahmed, N. H.; Chowdhary, A. // Indian Journal of Medical Microbiology;Apr2013, Vol. 31 Issue 2, p154 

    Purpose: The study was conducted to compare different methods of detection of pathogenic protozoan parasites in stool specimens of People Living with HIV/AIDS (PLHA). Materials and Methods: Stool specimens of 242 HIV sero-positive patients were examined using the wet mount technique, modified...

  • Evaluation of in vivo and in vitro Biological Activity of a Vibrio Cholerae 01 Hemolysin. Galindo, José Arellano; Angeles, Maria Guadalupe Rodriguez; Guadarrama, Norma Velazquez; Esteban, Enrique Santos; Cerezo, Silvia Giono // Clinical & Investigative Medicine;Dec2007, Vol. 30 Issue 6, pE250 

    Purpose: To evaluate the hemolysin effect by ileal loop model produced by Vibrio cholerae O1 strains, compared with the cellular lysis or cytotoxic activity (CA) observed in cell culture. Method: We studied nine V. cholerae O1 strains, obtained during the Mexican outbreak of cholera (1990-1993),...

  • A phase I pharmacokinetic and pharmacodynamic study of BGC9331 and carboplatin in relapsed gynaecological malignancies. Benepal, T.; Jackman, A.; Pyle, L.; Bate, S.; Hardcastle, A.; Aherne, W.; Mitchell, F.; Simmons, L.; Ruddle, R.; Raynaud, F.; Gore, M. // British Journal of Cancer;10/17/2005, Vol. 93 Issue 8, p868 

    BGC9331 is a rationally designed, specific nonpolyglutamatable thymidylate synthase (TS) inhibitor that is active in gynaecological malignancies. In the light of the sensitivity of human ovarian tumour cell lines to BGC9331 and non-cross resistance to platinum drugs, we studied the combination...

Share

Read the Article

Courtesy of THE LIBRARY OF VIRGINIA

Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics