Method for preparing a K serum with a vibrio parahaemolyticus as an antigen

Patent metadata

Field	Value
Publication number	US-10994005-B2
Application number	US-201916771693-A
Country	US
Kind code	B2
Filing date	Jul 30, 2019
Priority date	May 30, 2019
Publication date	May 4, 2021
Grant date	May 4, 2021

Abstract

A novel K (capsular antigen) serotype of Vibrio parahaemolyticus and an application thereof are provided. A novel K (capsular antigen) serotype of Vibrio parahaemolyticus, which was deposited at the China General Microbiological Culture Collection Center (Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing) on Feb. 20, 2019, with a deposit number of CGMCC No. 17249, wherein the K epitope of the Vibrio parahaemolyticus has a sequence set forth in Sequence No. 1. The novel K serum is highly specific, and can be used to conveniently and quickly detect a novel K serotype of Vibrio parahaemolyticus (O4:KUT-recAin) which has a rising infection rate in recent years. It provides important detection techniques for the pathogen diagnosis, monitoring and prevention of infectious diarrhea.

First claim

What is claimed is:

1. A method for preparing a K serum with a Vibrio parahaemolyticus as an antigen, comprising:
the Vibrio parahaemolyticus is seeded into a BHI medium with 3% NaCl, and cultured at 37° C. overnight;
the bacteria are collected, washed twice with normal saline, and then inactivated with a normal saline solution containing 0.5% formaldehyde for 24 h, and the inactivated bacteria are then dissolved in normal saline to act as an antigen; and
the K serum is prepared by injecting and immunizing an animal with the Vibrio parahaemolyticus as the antigen;
wherein the Vibrio parahaemolyticus was deposited at the China General Microbiological Culture Collection Center (Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing) on Feb. 20, 2019, with a deposit number of CGMCC No. 17249, wherein the K epitope of the Vibrio parahaemolyticus has a sequence set forth in Sequence No. 1.

2. The method for preparing the K serum with a Vibrio parahaemolyticus as an antigen according to claim 1 , further comprising:
healthy 12-week-old New Zealand white rabbits are selected and immunized with the inactivated Vibrio parahaemolyticus in 0.85% NaCl at a concentration of 109 CFU/ml, 4 sites for each rabbit and 250 μl for each site, and subsequently boosted once every 2 weeks in an immunization volume the same as the first immunization, 5 times in total;
thereafter, whole blood is collected from the heart, and the collected whole blood is left at 37° C. for 2 h, transferred to 4° C. overnight, and then centrifuged to collect serum, which is the K serum.

Assignees

• Univ Zhejiang

Inventors

• Chen Yu
• Chen Xiao
• Zhu Qiaoyun
• Xie Guoliang
• Wang Ruonan

Classifications

• C07K2317/21

  from primates, e.g. man · CPC title

• C12N1/205

  Bacterial isolates · CPC title

• C07K16/065

  Purification, fragmentation · CPC title

• A61K2039/521

  inactivated (killed) · CPC title

• A61K39/107 Primary

  Vibrio · CPC title