Gene detecting film strip for multiple bacteria and primer thereof

Abstract

The invention discloses a gene detecting film strip for multiple bacteria and a primer thereof. Ten common bacteria are selected as detected objects and 16SrRNA (Ribosomal Ribonucleic Acid) is used as a detection target gene; and a detection probe of the ten common bacteria and a matched primer thereof are designed. A cellulose nitrate thin film strip is used as a substrate for fixing the probe and used for preparing a gene detecting film strip of the ten common bacteria. The 16SrRNA gene segment of the bacteria and a biotin mark are subjected to PCR (Polymerase Chain Reaction) amplification.After the PCR amplified segment is hybridized with the film strip, the biotin mark is detected by streptavidin marked by alkaline phosphatase and then an NBT (Nitroblue Tetrazolium) / BCIP (5-bromo-4-chloro-3'-indolyphosphate P-toluidine salt) color development is carried out. The result shows that the film strip can independently detect any one or more of the ten bacteria. The invention successfully researches the gene detecting film strip for the ten common bacteria and establishes a method for detecting bacteria by the film strip; and the method provided by the invention has the advantages of high throughput, high speed, accuracy and lower cost and has a good clinical application prospect.

Description

technical field [0001] The invention belongs to the field of gene detection, and in particular relates to a gene detection membrane strip and a primer thereof which can be used for various bacteria. Background technique [0002] Infectious and infectious diseases caused by pathogenic bacteria are important factors that threaten human health and cause social panic. Rapid and accurate detection of pathogenic bacteria is the key to effective prevention and treatment of infectious diseases. Conventional physiological, biochemical and serological identification methods are cumbersome, long-term, and difficult to cultivate some pathogenic bacteria. Although PCR, ELISA and other methods shorten the detection cycle, the false positive rate is high [1，2] ,. [0003] The rise of gene chip technology has made it possible to detect pathogenic bacteria quickly, in small amounts, accurately and with high throughput [3] . The target genes selected for gene chip detection of pathogenic ...

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Problems solved by technology

However, fluorescently labeled gene chips require expensive instruments such as laser confocal microarray scanners and gene chip spotters. [6] , high cost, expensive

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