Application Of Colloidal Gold Immunochromatography Detected Monocytogenes Listeria Monocytogenes

Background and Purpose：In recent years, the infection caused by Listeria monocytogenes infood-borne diseases are common in China and other countries of theworld, there are about6-12people for every100,000people in theclinical morbidity of United States and Europe and other developedWestern countries, the mortality rate of20%-30%or higher[4]. Clinicaldisease has been caused in some European and American countries by theListeria monocytogenes bacteria than Salmonella. The potential humanhealth hazards caused by bacteria are increasingly attracted worldattention. Therefore, rapid, sensitive and accurate detection of bacteria isan important means to clinically effective prevention and treatment oflisteriosis and to diagnose the pathogen[5].Method:Single increase in the traditional test method for Listeriamonocytogenes isolation and culture of the technical evaluation andsuspicious colonies take about1-2weeks, whether it is spent on thehuman or material resources are relatively large[6,7], and detection rate isrelatively low and not suitable for application in the actual inspection work. PCR method for detection of bacteria with rapid, specific, sensitiveand high, but because of DNA degradation in dead bacteria is slow, thismethod for the distinction between viable and dead bacteria is not veryclear, easy to produce false-positive[8,9]; So first in the clinicalrequirements for detection of Listeria monocytogenes specimens werecultured, so you can minimize the probability of false-positive, butwhen the content of many of the dead bacteria in the specimens andinstrumental analysis for a long time and inevitably emergent thefalse-positive signal. False-positive detection of the clinical treatment of awrong information and judgment will be not only economic loss willresult in serious consequences. To establish a simple, sensitive, rapid andspecific detection of Listeria monocytogenes is necessary.90years of the20th century, developed the colloidal goldimmunochromatography technique is chromatography and immunogoldmetallographic combination of diagnostic techniques, the technologyafter years of use, a number of advantages: fast, easy to operate, andELISA similar specificity and sensitivity of the advantages[10], can bewidely used in biological, medical and other research fields, especially inthe more common medical tests. In this study, using colloidalgold-labeled single growth of Listeria monocytogenes in antibodypreparation immunochromatographic detection reagent plate, withintuitive, specificity and sensitivity characteristics of L. monocytogenes tested.Results:(1) Filter out the desired monoclonal antibodies by hybridomatechnology.(2) Sodium citrate reduction of visible light absorbance of theprepared colloidal gold, analyse colloidal gold particles size anddistribution,that the maximum absorption peak wavelength of525nm,20nm diameter colloidal gold.(3)Colloidal gold to detect single Listeria bacteria Yin, positiveresults, detection limit is105cfu/mL.Conclusion:(1) used in this study produced a single nuclear Listeriamonocytogenes bacterial protein immunized animals Balb/c mice, thepreparation of hybridoma cell lines, preparation of ascites antibody. Pureantibody titer, compared with the previous method to improve thesensitivity and accuracy.(2)Colloidal gold-labeled single increase in the Preparation ofimmunochromatographic detection of Listeria monocytogenes antibodyreagents board, for detection of Listeria monocytogenes, has theadvantages of fast and convenient.