Identification method of ST37 type clostridium difficile and unit point variation type ST81 thereof

A Clostridium difficile and identification method technology, which is applied in the field of strain type identification, can solve the problems of cumbersome operation of the identification method, inability to use conventional detection, and long time-consuming, etc., and achieves easy promotion and application, simple method, and easy operation Effect

Inactive Publication Date: 2017-09-26
THE SECOND HOSPITAL OF HEBEI MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide an identification method for ST37 Clostridium difficile and its single-site variant ST81, so as to solve the problems that the existing identification methods are cumbersome, time-consuming and unable to be used for routine detection

Method used

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  • Identification method of ST37 type clostridium difficile and unit point variation type ST81 thereof
  • Identification method of ST37 type clostridium difficile and unit point variation type ST81 thereof
  • Identification method of ST37 type clostridium difficile and unit point variation type ST81 thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Samples: 159 strains of Clostridium difficile were obtained from the Hebei Medical Microbial Culture Collection Center (HBCMCC), among which 152 strains of Clostridium difficile were randomly selected from diarrhea patients and healthy population isolates collected from 2010 to 2014 by using the method of random number table (65 strains were collected from patients with diarrhea, 87 strains were collected from healthy adults or children), and 7 strains of Clostridium difficile standard strains (ATCC BAA-1870, ATCC BAA-1382, ATCC BAA-1812, ATCC BAA-1812, ATCC BAA-1804, ATCC9689, ATCC 43255 and ATCC 43598).

[0023] The strains were stored in the HBCMCC ultra-low temperature refrigerator, and the strains were resuscitated before use. The strains were inoculated on Columbia blood agar plates (OXOID, UK), and cultured in an anaerobic environment at 37°C for 48 hours. After 48 hours, a single colony with typical morphology was scraped for The second passage, cultured under t...

Embodiment 2

[0043] Samples: Randomly select 3 strains of Clostridium difficile ST35, 4 strains of Clostridium difficile ST2 and 3 strains of ST3 difficile, 1 C. difficile ST8, 4 C. difficile ST54, 1 C. difficile ST139, 1 C. difficile ST81 and 22 C. difficile ST37, all from HBCMCC .

[0044] Reagents used: with embodiment 1.

[0045] Instruments used: VITEK MS automatic rapid microbial mass spectrometry detection system, BioMérieux, France.

[0046] Each sample was tested as follows.

[0047] MALDI-TOF sample preparation: the method is the same as in Example 1.

[0048] data collection:

[0049] VITEK MS: Same as Example 1.

[0050] Data analysis: Detect whether there are protein peaks at m / z3235-3245 and m / z3280-3290 in the mass spectrum, or export a TXT file containing the peak list, and observe whether there are protein peaks at m / z3235-3245 and m / z3280-3290.

[0051] The identification results are shown in Table 1. The test results of 39 strains from different sources in the tabl...

Embodiment 3

[0057] Samples: 2 strains of Clostridium difficile ST37 (numbered 38 and 90 respectively) and 1 Clostridium difficile ST81 (numbered 81) were randomly selected from the strains of Clostridium difficile collected in different years (2010, 2012, 2014). ), 1 strain of Clostridium difficile ST3 (No. 3), 1 strain of Clostridium difficile ST102 (No. 45) and 1 strain of Clostridium difficile ST348 (No. 86), a total of 6 strains, all of which were from HBCMCC. See Table 2 for the strain number, type and year of collection.

[0058] Table 2 strain information

[0059]

[0060] The 6 strains of Clostridium difficile were inoculated and passaged twice at different times by three experimenters respectively, and the culture conditions were anaerobic culture at 37°C for 48 hours (anaerobic culture system, Guangzhou Youde Company). Randomly select three colonies in the medium of the second passage of each strain, and collect a total of 54 Clostridium difficile colonies as identification ...

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Abstract

The invention provides an identification method of ST37 type clostridium difficile and unit point variation type ST81 thereof. The identification method comprises the following steps: protein in a clostridium difficile strain to be identified is extracted to obtain a mass spectrometric text sample; and mass spectrometric detection equipment is utilized to detect the sample, and strains with specific protein peaks at m / z 3235-3245 and m / z 3280-3290 in an obtained mass spectrogram are the ST37 type clostridium difficile or the unit point variation type ST81 thereof. The invention establishes an MALDI-TOF MS-based rapid identification method of the ST37 type clostridium difficile and the unit point variation type ST81 thereof for the first time. The method has the characteristics of real time and rapidness, can screen the ST37 type clostridium difficile and the unit point variation type ST81 thereof in real time while identifying the strains, only needs to add 1min for the detection process of ST37 or ST81 type on the basis of strain identification without other special instruments or equipment or additional operation steps and has important significance on rapidly diagnosing diseases and timely providing clinical medication guidance.

Description

technical field [0001] The invention relates to an identification method of bacterial strains, in particular to an identification method of ST37 type Clostridium difficile and its single point variant ST81. Background technique [0002] Clostridium difficile (CD) is an obligate anaerobic, gram-positive Bacillus macrospores, the main pathogen of antibiotic-associated diarrhea and pseudomembranous colitis (PMC), accounting for about 30% of antibiotic-associated diarrhea And more than 90% of pseudomembranous colitis is caused by Clostridium difficile. The clinical manifestations of Clostridium difficile-associated diarrhea (CDAD) are quite different, and the symptoms of patients can be mild diarrhea or severe fatal pseudomembranous colitis. The genotype of Clostridium difficile is related to the patient's condition, and the development and prognosis of the disease may be completely different if infected with different genotypes, such as infection with a highly virulent genotyp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/62
CPCG01N33/6851
Inventor 赵建宏李茹鑫杨靖孙素菊李志荣强翠欣魏宏莲时东彦翟宇王晓明赵杏珍赵宝鑫赵峰
Owner THE SECOND HOSPITAL OF HEBEI MEDICAL UNIV
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