Tracing method of cronobacter spp. in infant formula milk powder

A technology of Cronobacter and infant formula, which is applied in the field of traceability of Cronobacter in infant formula milk powder, can solve problems such as restricting the healthy development of the PIF industry, inability to prevent and control Cronobacter, and the impact of pathogenic bacteria

Inactive Publication Date: 2016-09-21
NORTHEAST AGRICULTURAL UNIVERSITY
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Problems solved by technology

Cronobacter is the species with the strongest environmental adaptability in Enterobacteriaceae, so although the processing environment of PIF (including raw materials, pretreatment, spray drying, fluidized bed and packaging, etc.) has been strictly regulated and However, the contamination rate of Cronobacter in commercially available PIF is still as high as 4.3%. It can be seen that the pollution of this pathogen

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  • Tracing method of cronobacter spp. in infant formula milk powder
  • Tracing method of cronobacter spp. in infant formula milk powder
  • Tracing method of cronobacter spp. in infant formula milk powder

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Experimental program
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Effect test

Embodiment 1

[0033] Activation, purification and identification of embodiment 1 Cronobacter

[0034] The tested strains were 80 strains of Cronobacter, of which 43 were isolated from PIF, 27 were isolated from the processing environment of PIF, and 4 were reference strains of Cronobacter (C. sakazakii ATCC BAA-894, C. sakazakii ATCC 29004 , C.sakazakii ATCC 29544 and C.sakazakii ATCC 12868), 6 strains were Cronobacter type strains (C.malonaticus CDC105877T, C.dublinensis LMG23823T, C.turicensis LMG23827T, C.universalis NCTC9529T, C.condimenti LMGT and C. .muytjensii ATCC 51329T). In addition, E.aerogenes ATCC 13048, E.cloacae ATCC 35030, Escherichia CMCCB 44113 and Escherichia O157 were used as external reference strains. The specific conditions are shown in Table 4, and the above-mentioned tested strains were all from Laboratory 503, Key Laboratory of Dairy Science, Ministry of Education, Northeast Agricultural University.

[0035] Table 4 Tested strains and their sources

[0036] Tabl...

Embodiment 2

[0050] The multi-locus sequence typing of embodiment 2 Cronobacter

[0051] (1) 7 pairs of housekeeping genes specific PCR amplification and gene sequencing

[0052] (1) Specific amplification of 7 pairs of housekeeping genes

[0053] Synthesize 7 pairs of primers for housekeeping genes (atpD, fusA, glnS, gltB, gyrB, infB, and ppsA) (see Table 3 for the gene sequence), use Cronobacter DNA as a template for specific amplification, and PCR of 7 pairs of primers The amplification system and conditions are the same, as shown in Table 6. The PCR amplification product was checked by 1% agarose gel electrophoresis, and the results were observed and recorded under the UVP ultraviolet gel phase system. The results are as follows: figure 1 shown. The target bands of the 7 housekeeping genes were clear, and the size of the product sequence was judged to be the target bands of the 7 housekeeping genes according to the position of the bands.

[0054] Table 6 7 pairs of housekeeping gen...

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Abstract

The invention provides a tracing method of cronobacter spp. in infant formula milk powder. The method comprises the steps that by means of a multiple-locus sequence typing technology, molecular typing research is conducted on the cronobacter spp. of PIF raw materials, semi-finished products, finished products and processing environments thereof, population characteristics of the cronobacter spp. are revealed, a phylogenetic relationship of the cronobacter spp. is confirmed, phylogenetic analysis is conducted on the cronobacter spp. through bioinformatics, and therefore genetic characteristics of the cronobacter spp. in the above-mentioned environments are revealed systematically; on the basis of cronobacter spp. MLST typing, correlation analysis is conducted by taking ST of pathogenic bacteria and environmental sources as variables, key links which are most prone to pollute the cronobacter spp. in the PIF processing process and distribution rules are determined, and therefore the source of the cronobacter spp. in the infant formula milk powder is traced. The purpose of fundamentally monitoring the PIF production process and ensuring the PIF quality are achieved.

Description

technical field [0001] The invention belongs to the field of food safety, and particularly relates to a method for tracing the source of Cronobacter in infant formula milk powder. Background technique [0002] Cronobacter spp. is one of the most harmful opportunistic pathogens in infant formula (powdered infant formula, PIF). PIF contaminated by Cronobacter can lead to severe necrotizing enterocolitis, bacteremia and meningitis and other malignant diseases, with a mortality rate as high as 40%-80%. Cronobacter is the species with the strongest environmental adaptability in Enterobacteriaceae, so although the processing environment of PIF (including raw materials, pretreatment, spray drying, fluidized bed and packaging, etc.) has been strictly regulated and However, the contamination rate of Cronobacter in commercially available PIF is still as high as 4.3%. It can be seen that the pollution of this pathogenic bacteria has seriously affected and restricted the healthy develop...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6869C12Q1/689
Inventor 姜毓君满朝新费鹏孙露宏潘瑞丽
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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