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Marker for diagnosing drug resistance of streptococcus pneumoniae to vancomycin and application of marker

A technology of Streptococcus pneumoniae and vancomycin, applied in the direction of microorganisms, microorganism-based methods, biochemical equipment and methods, etc., can solve problems such as time-consuming, limited space, and large volume

Active Publication Date: 2021-04-13
遵义医科大学珠海校区
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantages of the prior art mainly include: (1) it takes a long time, and it needs to be established on the basis of separation and culture for drug sensitivity detection, which needs 4-8 weeks for separation and culture, and on this basis, the follow-up traditional solid ratio method (4- 8 weeks) or liquid drug susceptibility (1-2 weeks); (2) due to site constraints, a larger incubator is required, and a fixed culture room is required to ensure biosafety

Method used

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  • Marker for diagnosing drug resistance of streptococcus pneumoniae to vancomycin and application of marker
  • Marker for diagnosing drug resistance of streptococcus pneumoniae to vancomycin and application of marker
  • Marker for diagnosing drug resistance of streptococcus pneumoniae to vancomycin and application of marker

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Experimental program
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Embodiment 1

[0039] In the present invention, vancomycin-sensitive Streptococcus pneumoniae VSSP (or wild-type Streptococcus pneumoniae) is domesticated and treated with vancomycin to obtain the vancomycin-resistant Streptococcus pneumoniae VRSP, and then iTRAQ differential proteomics is used to compare and analyze VSSP and VRSP A total of 260 differentially expressed proteins (DEPs) with a differential fold greater than or equal to 2.0 and p<0.01 were identified in three independent replicates, of which 129 DEPs were up-regulated in VRSP and 131 DEPs were up-regulated in VRSP. expression decreased.

[0040] In order to further screen out the proteins potentially regulating VRSP formation, we performed KEGG pathway enrichment analysis on DEPs, and used cytoscape software to construct the interaction map between these DEPs and pathways, such as figure 1 As shown, the two-component system regulatory protein CiaR was significantly overexpressed in VRSP, while the two-component system ComD and...

Embodiment 2

[0042] To further verify the reliability of mass spectrometry data, we used RT-qPCR to detect the expression of ciaR, comD and comE in VRSP. like figure 2 Panel A shows that ciaR was significantly up-regulated in VRSP compared with VSSP, while comD and comE were significantly down-regulated in VRSP, consistent with the proteomic results.

[0043]It is known that the two-component regulatory systems CiaRH and ComDE are the key proteins regulating the competence formation of Streptococcus pneumoniae, and CiaRH is negatively correlated with competence, and ComDE is positively correlated with competence. Since the ability of Streptococcus pneumoniae to take up exogenous DNA (ie, transformation efficiency) is positively correlated with the formation of competence, the ability of VSSP and VRSP to take up exogenous DNA was compared and analyzed. like figure 2 As shown in panel B, whether it is plasmid DNA or PCR fragment DNA, the ability of VRSP to take up exogenous DNA is signif...

Embodiment 3

[0045] In order to further study the relationship between the two-component system response regulatory protein CiaR and Streptococcus pneumoniae resistance to vancomycin, the present invention constructed a ciaR overexpression strain (ciaR OE) and a ciaR deletion mutant (ciaR mutant), and used the time sterilization curve. The vancomycin tolerance of wild-type strains, ciaR overexpression strains and ciaR deletion mutants was compared and analyzed.

[0046] The result is as image 3 As shown in A, the present invention has successfully constructed a ciaR overexpression strain and a ciaR deletion mutant strain. like image 3 As shown in middle B, the same amount of wild-type strains, ciaR overexpression strains and ciaR deletion mutant strains were then taken and treated with 10×MIC vancomycin respectively, and the plates were sampled and counted at different time points. The survival rate of the expression strain was about 20 times higher than that of the wild type strain, w...

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Abstract

The invention belongs to the field of biological detection, and discloses a marker for diagnosing drug resistance of streptococcus pneumoniae to vancomycin and application of the marker. The marker for diagnosing the drug resistance of streptococcus pneumoniae to vancomycin is composed of ciaR, comD and comE. And when at least one of comD or comE is lower than the expression quantity of the wild type streptococcus pneumoniae strain and the expression quantity of ciaR is higher than the expression quantity of the wild type streptococcus pneumoniae strain, it judges that streptococcus pneumoniae is resistant to vancomycin. The marker can be used for detecting the tolerance degree of streptococcus pneumoniae to vancomycin. Based on the marker, a corresponding product for detecting the drug resistance of streptococcus pneumoniae to vancomycin can be prepared and plays an important role in coping with the drug resistance of streptococcus pneumoniae.

Description

technical field [0001] The invention belongs to the field of biological detection, and particularly relates to a marker for diagnosing the resistance of Streptococcus pneumoniae to vancomycin and its application. Background technique [0002] Streptococcus pneumoniae is the most common pathogen causing community-acquired pneumonia. It often resides in the upper respiratory tract and can cause a series of diseases such as pneumonia, otitis media, meningitis, bacteremia and sepsis, resulting in 1-2 million infant deaths each year. Antibiotics are the main means of treating Streptococcus pneumoniae infection. In the past, penicillin could kill Streptococcus pneumoniae; but with the extensive use of antibiotics, penicillin-resistant Streptococcus pneumoniae (PRSP) has spread worldwide (with the multidrug resistance to other antibiotics). Strains in some regions are resistant to penicillin as high as 45%, making the treatment of Streptococcus pneumoniae infections increasingly d...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/6851C12N15/11C12Q1/14C12R1/46
CPCC12Q1/689C12Q1/6851C12Q2600/106C12Q2600/158C12Q2600/166C12Q2531/113C12Q2545/101
Inventor 阳小燕乐尧金
Owner 遵义医科大学珠海校区