A PCR primer and method capable of detecting Salmonella and identifying Salmonella pullorum

Abstract

The invention discloses a PCR primer and a method capable of detecting Salmonella and identifying Salmonella pullorum. According to the Salmonella target sequence (SEQ ID NO.1, NO.2, NO.3), a pair of specific primers were screened and designed, including primer SEEP‑F and primer SEEP‑R, wherein the nucleotide sequence of SEEP‑F As shown in SEQ ID NO.7; the nucleotide sequence of SEEP‑R is shown in SEQ ID NO.8. The method is to use the above-mentioned primers to perform PCR amplification on the sample to be tested to obtain PCR amplification products; use agarose gel electrophoresis or sequencing to detect the amplification products; Salmonella pullorum; if the size of the amplified product is 1394bp or 1400bp, the sample contains or is candidate to contain other Salmonella. The invention can quickly detect Salmonella and identify Salmonella pullorum, and is quicker, simpler and lower in cost compared with traditional salmonella cultivation, biochemical identification and Salmonella pullorum identification methods.

Description

technical field [0001] The invention belongs to the technical field of PCR detection, and in particular relates to a PCR primer and a method capable of detecting Salmonella and identifying Salmonella pullorum. Background technique [0002] Salmonella is a zoonotic pathogen with important public health significance, and more than 2500 serotypes have been reported so far. Salmonella pullorum can cause pullorum, which has a high mortality rate for chicks and brings serious economic losses to large-scale chicken farms. Salmonella pullorum is mainly prevalent in chickens aged 2-3 weeks, but at present there is a trend of increasing susceptible days and diversifying clinical disease types, resulting in the whole chicken raising cycle being harmed by this disease. Chickens with pullorum disease often show drowsiness, weakness and loss of appetite, gather together, droop their wings, make sharp calls when defecating, and often have pinkish white or green excrement around the anus, ...

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Problems solved by technology

Among them, "Salmonella and its five serotypes multiplex PCR identification kit" (authorization number CN201310110380), "primer set, kit and method for identifying Salmonella pullorum and Salmonella gallinarum typhi" (application number 201510570362.9) involved pullorum Salmonella detection primers can also amplify multiple Salmonella serotypes such as Wandsworth, Manchester, Chester, Java, Bareilly, etc., with poor specificity; "PCR detection primers and detection methods for Salmonella pullorum" (application number 201410492056. The gene ipaJ for the detection of Salmonella pullorum selected in "PCR Detection Kit for Identification of Pullorum Pullorum" (application number 201611111312.5) also exists in Salmonella pullorum, and the gene is located on a small plasmid, not all Salmonella pullorum carry it, The situation of missed detection is prone to occur; while "A PCR detection kit for rapid identification of specific serotype Salmonella" (application number 201610459944.4), "A PCR detection kit for rapid identification of pullorum and Salmonella gallinarum typhi" (201610628075.3 The tcpS and flhB genes selected in ) are not specific to Salmonella pullorum, and the detection results cannot accurately determine the pullorum serotype

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