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Composition and kit for detection and analysis of strains of clostridium difficile and method of detecting strains of clostridium difficile by using the same

一种艰难梭菌、组合物的技术，应用在基于微生物的方法、生物化学设备和方法、微生物等方向，能够解决变异假阳性测定、错误等问题

Inactive Publication Date: 2014-01-01
SAMSUNG ELECTRONICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In using these virulence genes for molecular diagnostics, there is the possibility of errors due to genetic variation and the possibility of false positive assays due to variation

Method used

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  • Composition and kit for detection and analysis of strains of clostridium difficile and method of detecting strains of clostridium difficile by using the same
  • Composition and kit for detection and analysis of strains of clostridium difficile and method of detecting strains of clostridium difficile by using the same
  • Composition and kit for detection and analysis of strains of clostridium difficile and method of detecting strains of clostridium difficile by using the same

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Experimental program
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Effect test

Embodiment 1

[0062] Selection of primers and probes and PCR evaluation for detection of C. difficile

[0063] When a disease-causing gene is used for molecular diagnosis, there is the possibility of error occurring due to gene variation and the possibility of false positive determination. Thus, there is a need for molecular diagnostics to utilize genes with less variation and ubiquitous expression in various C. difficile strains.

[0064] gluD (glutamate dehydrogenase) is a protein that is ubiquitously expressed in various strains of C. Toxic is still poisonous.

[0065] Therefore, two primer sets for detecting Clostridium difficile strains and one probe for real-time PCR detection by specifically amplifying the gluD gene encoding gluD protein were designed through the NCBI website, and their interaction was analyzed and verified.

[0066] Primer sets and probes are as follows:

[0067] Forward primer: 5'-GCTGCATTAGAAAACTCTATAAC-3' (SEQ ID NO: 1),

[0068] Reverse primer: 5'-CAGCCTCT...

Embodiment 2

[0077] Primer and probe selection and PCR evaluation for the detection of toxin A in Clostridium difficile strains

[0078] Virulent C. difficile strains can be detected by toxin A, which induces C. difficile virulence.

[0079] A primer set for specifically amplifying tcdA (which is the gene of Clostridium difficile toxin A) and a probe for real-time PCR detection were designed via the NCBI website, and their interaction was analyzed and verified. Primer sets and probes designed for real-time PCR detection with improved sensitivity.

[0080] Primer sets and probes are as follows:

[0081] Forward primer: 5'-GCGGAGTATATTTAGATGTTG-3' (SEQ ID NO: 7),

[0082] Reverse primer: 5'-ACGGTCTAGTCCAATAGA-3' (SEQ ID NO: 8), and

[0083] Probe: 5'-ATGCTTCCAGGTATTCACT-3' (SEQ ID NO: 9).

[0084] Reference tcdA was amplified using the following primer set (see J Microbiol Methods 83:59-65 (2010)):

[0085] Forward primer: 5'-TTGTATGGATAGGTGGAGAAGTCAGT-3' (SEQ ID NO: 10), reverse prim...

Embodiment 3

[0091] Primer and probe selection and PCR evaluation for detection of Clostridium difficile toxin B

[0092] Virulent C. difficile can be detected by toxin B, which induces C. difficile virulence.

[0093] The toxin B gene tcdB is less conserved among C. difficile strains. A primer set for specifically amplifying the tcdB and a probe for real-time PCR detection were designed via the NCBI website, and their interactions were analyzed and verified.

[0094] Primer sets and probes designed for real-time PCR detection with improved sensitivity.

[0095] Primer sets and probes are as follows:

[0096] Forward primer: 5'-GGTGGTATGTATTTAGATGTTGA-3' (SEQ ID NO: 14),

[0097] Reverse primer: 5'-TCCACTGTTACTGAACTAGG-3' (SEQ ID NO: 15), and

[0098] Probe: 5'-CCAGGAATACAACCAGACT-3' (SEQ ID NO: 16).

[0099] The reference tcdB was amplified using the following primer set (see J Microbiol Methods 83:59-65 (2010)):

[0100] Forward primer: 5'-GAAACAGGATGGACACCAGGTT-3' (SEQ ID NO: 17...

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Abstract

A composition and kit for detecting Clostridium difficile including a primer set for detecting a strain of Clostridium difficile, and a method of detecting Clostridium difficile by using the same.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of Korean Patent Application No. 10-2012-0061671 filed with the Korean Intellectual Property Office on June 8, 2012, the disclosure of which is incorporated herein by reference in its entirety. technical field [0003] The present disclosure relates to compositions and kits comprising primer sets for the detection of Clostridium difficile strains or virulent Clostridium difficile strains, and detection of Clostridium difficile in a sample by using the compositions or kits. Clostridium strain method. Background technique [0004] Clostridium difficile is a species of anaerobic, spore-forming Clostridium gram-positive bacteria. [0005] C. difficile is the most serious cause of antibiotic-associated diarrhea and can cause pseudomembranous colitis, a serious colon infection that often results in the eradication of normal intestinal flora by antibiotics. C. difficile, which naturally r...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q2600/156C12Q1/689C12R2001/145
Inventor 郑善玉金俊镐李树官黄奎渊严泰喜
Owner SAMSUNG ELECTRONICS CO LTD
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