Quick detection method for newcastle disease virus and its immune colloidal gold test card

Abstract

The invention provides a method for quickly detecting Newcastle disease virus. The method comprises the following steps that: a Newcastle disease virus monoclonal antibody, a goat-anti-Newcastle disease virus polyclonal antibody and a goat-anti-Balb/C mouse immunoglobulin antibody are established; the treatment of a combination pad and NC membrane is completed; after a to-be-detected sample is added on a sample pad and filtered, the NDV in the sample is combined with the gold-labeled antibody in the combination pad; the gold-labeled antibody is firstly combined with the Ab2 on the NC membrane through chromatography to form a visible combining line, and an uncombined gold-labeled antibody undergoes chromatography continuously and is combined with Ab3 to form a second combining line; finally, a step of result observation, during which, two combining lines are formed when the result is positive, while one quality control line is formed when the result is negative. The method has the characteristics of simple operation, quickness, high sensitivity and excellent specificity, etc.; moreover, the method is low in cost and is suitable for Newcastle disease virus detection at grass root or clinic; in addition, the method can detect the Newcastle disease virus in bird blood and dejection and can obtain a result within 10 min with the sensitivity reaching to 10ng/ml. The invention also provides a Newcastle disease immune colloidal gold test card for realizing the method.

Description

technical field [0001] The invention relates to a technology for diagnosing or detecting Newcastle disease virus in farms, epidemic prevention and quarantine departments, medical institutions and other units, in particular to a method for quickly detecting Newcastle disease virus and a colloidal gold test card for Newcastle disease virus immunity. Background technique [0002] Newcastle disease virus (ND) is an acute, septic, and highly contagious infectious disease, which spreads rapidly and has high morbidity and mortality. Once an outbreak occurs, it will cause immeasurable economic losses to the chicken industry. Therefore, farms, epidemic prevention and quarantine departments, and medical institutions often need diagnosis or test results. The General Administration of Quality Supervision, Inspection and Quarantine issued a document in 2001 requiring regular monitoring of meat export enterprises every two months. This regulation requires the use of chicken embryo virus ...

AI Technical Summary

Problems solved by technology

The chicken embryo virus isolation method is a sensitive and specific method, but it takes a long time, about 3 weeks, and the operation is cumbersome, it can only be used for monitoring, not for rapid detection, which is not conducive to rapid customs clearance and control of virus carried by poultry meat

CN1827775A (patent application number 200510008997.6) "Nucleoside sequence, kit and test method for detection of Newcastle virus", the invention selects Newcastle disease virus F gene (synthetic F protein) as the target region, and the selection can reflect the characteristics of the cleavage site and lack Design multiple pairs of primers and probes for the conserved region of the secondary structure, and establish fluorescent PCR to detect Newcastle disease virus. This method is 1,001,000 times more sensitive than the traditional PCR method, but it takes about 4 hours from sample processing to the result. The implementation of this method It also needs expensive instruments such as fluorescent quantitative PCR instrument, which is not suitable for promotion at the grassroots level

However, the required equipment is expensive and has been greatly limited in clinical testing

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