Enterobacter sakazaii enriched reagent and its production method

A technology for enriching Enterobacter sakazakii and Enterobacter sakazakii, applied to measuring devices, instruments, scientific instruments, etc., to achieve wide application prospects, simple separation, and improved sensitivity and detection rate

Inactive Publication Date: 2009-07-15
GUANGDONG INST OF MICROORGANISM
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved in the present invention is to provide a reagent for enriching Enterobacter sakazakii, which can directly enrich the target microorganisms from the sample or the pre-enrichment culture solution to play a concentration role and solve the problem of sample pretreatment. Bottleneck problem, improved detection sensitivity and detection rate

Method used

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  • Enterobacter sakazaii enriched reagent and its production method
  • Enterobacter sakazaii enriched reagent and its production method
  • Enterobacter sakazaii enriched reagent and its production method

Examples

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Embodiment 1

[0019] Example 1: Preparation of reagents for enrichment of Enterobacter sakazakii

[0020] 1. Preparation of Enterobacter sakazakii immune serum:

[0021] Enterobacter sakazakii immune serum containing polyclonal antibody against Enterobacter sakazakii was prepared according to conventional methods. The purified Enterobacter sakazakii were inactivated with 0.2% and 0.3% formaldehyde for 12h and 24h respectively, and 1ml was poured into TSA plates at fixed times for 24h at 37°C to observe the inactivation of the bacteria.

[0022] Take completely inactivated Enterobacter sakazakii, adjust the concentration to 108CFU / ml, take 0.1ml and emulsify it with an equal volume of Freund's complete adjuvant, and inject New Zealand rabbits subcutaneously at multiple points (in order to ensure the success rate, 3 rabbits were used at the same time) experiment), repeated injections every 2 weeks. One week after the last immunization (serum titer reached 1:50000), the rabbits were bled to ...

Embodiment 2

[0039] 1. Immunomagnetic beads were used to enrich Enterobacter sakazakii after enrichment of milk powder samples

[0040] Use the reagent for enriching Enterobacter sakazakii described in Example 1 (Enterobacter sakazakii immunomagnetic beads) to enrich the milk powder sample enrichment solution, take 100 μl, dilute or not, apply a chromogenic medium plate, and incubate at 37°C Then observe the typical blue-green colonies. On the chromogenic medium, the visible colonies are clear and dispersed, easy to observe and identify after separation and purification.

[0041] 2. Enrichment of Enterobacter sakazakii immunomagnetic beads combined with chromogenic medium for detection of Enterobacter sakazakii in milk powder

[0042] After enrichment of 37 suspected positive milk powder samples, the established immunomagnetic beads were used for enrichment, and the suspension was coated on a chromogenic medium plate of Enterobacter sakazakii. As a result, 11 samples were positive and ide...

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Abstract

The invention discloses an Enterobacter sakazakii enrichment reagent and preparation method thereof, the reagent comprises immunomagnetic beads coated with polyclonal antibody of Enterobacter sakazakii. The reagent concentrates the object micro-organism by directly enriching the object micro-organism in sample or bacterial enrichment culture medium and the key problem of the sample pre-processing is solved and the detection sensitivity and detection rate can be improved.

Description

technical field [0001] The invention belongs to the field of food microbe safety monitoring, and in particular relates to a reagent for enriching Enterobacter sakazakii and a preparation method thereof. Background technique [0002] Enterobacter sakazakii (Enterobacter sakazakii) is a pathogenic bacteria in milk powder that has attracted widespread attention in recent years. Enterobacter sakazakii, formerly known as Enterobacter cloacae yellow, was not considered a new strain until 1980, and was named after Japanese microbiologist Riichi Sakazakii, Enterobacter sakazakii. In 2008, the Codex Committee on Food Hygiene agreed to consider creating a new genus Cronobacter, which includes Enterobacter sakazakii and related species. Enterobacter sakazakii is a Gram-negative pathogenic bacterium parasitic in the intestinal tract of humans and animals. [0003] The source of E. sakazakii contamination in nature is still unclear (Friedemann et al., 2007; Lin et al., 2007), and E. sa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/531
Inventor 吴清平周艳红徐晓可张菊梅
Owner GUANGDONG INST OF MICROORGANISM
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