Reagent kit capable of simultaneously detecting monocytosis Listeria monocytogenes, bacillus cereus, cronobactersakazakii and staphylococcus aureus

Abstract

The invention discloses a one-tube multi-amplification internal-control PCR (polymerase chain reaction) reagent kit capable of simultaneously detecting monocytosis Listeria monocytogenes, bacillus cereus, cronobactersakazakii and staphylococcus aureus. The one-tube multi-amplification internal-control PCR reagent kit comprises a PCR premixed liquid and reference substances. A detection method includes acquiring samples; carrying out enrichment culture; extracting DNA (deoxyribonucleic acid) after enrichment culture is carried out; preparing PCR tubes; arranging the PCR tubes on PCR instruments and carrying out PCR amplification to obtain PCR amplification products; judging detection results of the PCR amplification products. The one-tube multi-amplification internal-control PCR reagent kit has the advantages that the monocytosis Listeria monocytogenes, the bacillus cereus, the cronobactersakazakii and the staphylococcus aureus can be simultaneously quickly detected by the aid of the one-tube multi-amplification internal-control PCR reagent kit by means of one-step reaction; the detection method is convenient to implement, good in reliability, short in detection period, high in sensitivity and specificity, low in cost and suitable for high-flux operation and standardized operation and includes simple operation steps as compared with existing detection methods; polymerase inhibiting factors and false negative results due to misoperation in sample treatment procedures can be indicated, accordingly, misjudgment on the results can be prevented, and the one-tube multi-amplification internal-control PCR reagent kit and the detection method are beneficial to improving the level of food safety detection and preventing and controlling foodborne diseases.

Description

technical field [0001] The present invention relates to biotechnology, in particular to a method capable of simultaneously detecting Listeria monocytogenes (hereinafter referred to as "Listeria monocytogenes"), Bacillus cereus, Cronobacter sakazakii, and Staphylococcus aureus. A multiplex system kit containing amplified internal standards for 4 food-borne pathogenic bacteria, which can simultaneously detect Listeria monocytogenes, Bacillus cereus, Cronobacter sakazakii, and Staphylococcus aureus in one reaction tube samples for identification testing. Background technique [0002] Foodborne disease covers a very wide range and is a serious public health problem all over the world, both in developed and developing countries. Surveillance data show that in 2011, one out of every 6.5 people in China suffered from foodborne diseases. The global incidence of foodborne illness is difficult to estimate, but a reported 1.8 million deaths were attributed to diarrheal diseases in 20...

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Problems solved by technology

Traditional biological methods take a long time and have complicated detection steps. Colloidal gold test strips are relatively expensive and have low detection sensitivity. Enzyme-linked immunosorbent assay (E

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