Preenrichment liquid culture medium for vibrio vulnificus clinical detection and application of preenrichment liquid culture medium

A technology of Vibrio vulnificus and bacteria enrichment solution, which is applied in the field of clinical pathogenic microorganism inspection, can solve the problems of ineffective inhibition of the growth of basophilic miscellaneous bacteria, missed detection of Vibrio vulnificus, and low specificity, so as to promote the development of Vibrio vulnificus. Bacterial growth, strong bacterial enrichment ability, and convenient preparation

Active Publication Date: 2015-11-25
THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the most commonly used enrichment solution is alkaline peptone water, which is often used for the selective enrichment culture of basophilic bacteria, especially Vibrio. The enrichment solution has a certain enrichment effect, but the composition is single and the specificity is not strong. , cannot effectively inhibit the growth of basophilic miscellaneous bacteria, and lack some carbon sources and growth factors that can promote the growth of Vibrio vulnificus. False negative results, resulting in missed detection of Vibrio vulnificus

Method used

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  • Preenrichment liquid culture medium for vibrio vulnificus clinical detection and application of preenrichment liquid culture medium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Isolation and identification of Clostridium butyricum WZMC1016 strain

[0043] The Clostridium butyricum WZMC1016 of the present invention has the following microbiological characteristics:

[0044] (1) Colony morphology: milky white, round and slightly convex, with irregular edges, 1 to 3 mm in diameter, slightly shiny on the surface, and bubbles in the liquid medium.

[0045] (2) Individual form: G + Bacillus, the bacteria are straight or curved rod-shaped, 0.6~1.2×3.0~7.0μm, round ends, single, pair, short chain, occasionally filamentous bacteria.

[0046] (3) Physiological and biochemical characteristics: maltose (+); mannitol (+); raffinose (+); lactose (+); ribose (+); starch (+).

[0047] Optimal conditions for cultivation: good growth under anaerobic conditions, but no growth under an aerobic environment. The optimum growth temperature is 35~40℃; the lowest growth temperature is 25~28℃; the highest is 43~45℃; the optimum growth pH is 6.5~7.0; the pH 4.5~5.0 or ...

Embodiment 2

[0050] The bacteria-enhancing liquid of the present invention is composed of the following weight materials:

[0051] Peptone 10g

[0052] Sodium chloride 30g

[0053] Growth factor 0.2g

[0054] Potassium chloride: 0.8g,

[0055] Magnesium chloride 2.5g,

[0056] 8g of sodium thiosulfate,

[0057] Growth factor 0.2g,

[0058] Polymyxin B400000IU, Polymyxin E100000IU,

[0059] Add distilled water to a total volume of 1000mL;

[0060] The proliferation factor is the broken product of Clostridium butyricum, and its preparation method and characteristic are that the process steps are:

[0061] (1) Fermentation of Clostridium butyricum: The seed solution of Clostridium butyricum WZMC016 strain was inserted into the sterilized sterilized peptone 1%, yeast extract 2%, glucose 2.5%, and ammonium sulfate 0.1% by weight. %, sodium bicarbonate 0.1%, manganese sulfate 0.02%, magnesium sulfate 0.02%, calcium chloride 0.002% liquid medium, after 36℃~38℃ anaerobic fermentation, 3000rpm 15-20min centrifuga...

Embodiment 3

[0069] The pre-enrichment solution used for rapid detection of Vibrio vulnificus is composed of the following substances:

[0070] Peptone 10g

[0071] Sodium chloride 30g

[0072] Magnesium chloride 3g

[0073] Potassium chloride 0.8g

[0074] Sodium thiosulfate 10g

[0075] Growth factor 0.2g

[0076] Add distilled water to a total volume of 1L;

[0077] Add polymyxin B40000IU and polymyxin E10000IU per 100mL of enrichment solution.

[0078] The preparation method and characteristics of the proliferation factor are the same as in Example 2;

[0079] The preparation process of the enrichment liquid is:

[0080] A. Weigh the above-mentioned weight and add distilled water to 1L, fully dissolve it, filter, and autoclave at 115°C for 20 minutes.

[0081] B. Add polymyxin B40000IU and polymyxin E10000IU to every 100ml of the sterilized enrichment solution obtained in step A, mix and dispense.

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Abstract

The invention relates to preenrichment fluid for vibrio vulnificus detection. A nitrogen source, a carbon source, inorganic salt, vitamins and other growth factors are added to the preenrichment fluid according to the biochemical features and culture features of vibrio vulnificus, and the preenrichment fluid has oxidation reduction potential, pH value and osmotic pressure suitable for the growth of the vibrio vulnificus. Compared with the alkaline peptone water widely used clinically and in food detection, the preenrichment fluid can facilitate the growth and resuscitation of the vibrio vulnificus in to-be-detected samples, can shorten culture time and increase positive detection rate, and can effectively inhibit growth and reproduction of other non-objected bacteria such as escherichia coli, pseudomonas and other gram positive bacteria. The preenrichment fluid is convenient to prepare, low in cost, high in enrichment ability and applicable to vibrio vulnificus detection of various levels of hospitals and detecting institutions.

Description

Technical field [0001] The invention belongs to the technical field of clinical pathogenic microorganism inspection, and specifically relates to a preparation method and application of a pre-enhancement solution for detecting Vibrio vulnificus. Background technique [0002] Vibriovulnificus (Vv) is an important food-borne pathogen. It is widely found in oceans, lakes, estuary water and seafood. Humans can also infect Vibrio vulnificus through raw seafood or wound contact with seawater with bacteria. This leads to severe systemic infections. The clinical symptoms mainly include fever, chills, vomiting, hypotensive shock, and characteristic bloody bullous lesions on the distal limbs. The process is quite rapid and the mortality rate is high. Lu Zhongqiu and others reported 34 cases of Vibrio vulnificus sepsis. The male to female ratio was 4.7:1. 76.5% of the patients had concurrent liver disease. The incidence was frequently from April to November, and the mortality rate was over 4...

Claims

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Application Information

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IPC IPC(8): C12Q1/04
CPCY02A50/30
Inventor 卢中秋李海啸洪广亮刘佳明
Owner THE FIRST AFFILIATED HOSPITAL OF WENZHOU MEDICAL UNIV
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