35 results about "Methicillin resistance gene" patented technology

The invention discloses a method for eliminating mecA plasmids based on CRISPR/Cas9 technology. The method comprises: selecting an MRSA strain to perform PCR amplification to a DNA sequence of a C terminal of mecA gene coding transpeptidase; carrying out gel extraction for mecA genes; connecting the mecA genes with a T-pMD19 (simple) carrier and preparing DH5[alpha] competent cells; electro-transforming the DH5[alpha] competent cells through the obtained T-pMD19-mecA plasmids; extracting the T-pMD19-mecA plasmids and performing sequencing and verification; performing double digestion treatment for the T-pMD19-mecA plasmids and performing gel extraction to the mecA genes; connecting pET-21a (+) plasmids with the mecA genes; designing and synthesizing oligos; constructing pCas9 :: mecA plasmids; electro-transforming the obtained pET-21a (+)-mecA plasmids into an escherichia coli expression strain BL21 (D3); and electro-transforming the pCas9 :: mecA plasmids and the pCas9 plasmids into BL21 (D3)+pET-21a(+)-mecA competent bacteria and BL21 (D3)+pET-21a(+) competent bacteria. The method is simple to operate and good in specificity, and can effectively block spread of mecA to eliminating the mecA strain.

Disclosed are methods of identifying a methicillin-resistant Staphylococcus aureus (MRSA) or methicillin-sensitive Staphylococcus aureus (MSSA) in a sample. The present invention provides a diagnostic method comprising modification of sequences of S. aureus by converting non-methylated cytosine residues ultimately into thymidine residues in the target nucleic acid. The invention further provides for the detection of modified sequences derived from the spa gene, the mecA gene, and the integrated SCCmec cassette of S. aureus.

The invention relates to a double-chain small molecule interference nucleic acid for inhibiting and killing various drug tolerance bacteria using methicillin-resistant staphylococcus aurous as represents. The siNA of the invention is a double-chain molecule with 19 base pairs. A sense strand and a antisense strand respectively have two overhanging bases dT at 5' terminals, the GC content is 40-55; the aimed target sequence is selected from the genes relative with the vital movements of copy, transcription and translation in the staphylococcus aurous genome and an mecA gene correlative with the drug tolerance; said target sequence is preserved in 900f staphylococcus aurous; said target sequence is in a conservative sequence area in more than 900f staphylococcus aurous with distinct source with all the gene sequences in the human genome; the target sequence of the said siNA double-chain molecule is selected from SEQ ID NO. 1-325, the sense strand is a corresponding DNA or RNA sequence with the target sequence and the antisense strand is a corresponding RNA or DNA with the sense strand according to the complementary base law.

The invention discloses a reagent kit and method for quickly detecting staphylococcus MecA. The reagent kit comprises cell lysate, amplifying reaction liquid, colloidal gold detection liquid and an immunochromatography reagent strip. The reagent kit is characterized in that transcription products of MecA and 16S rRNA are obtained through NASBA amplification on a primer pair in the amplifying reaction liquid. The reagent kit for quickly detecting genes of the staphylococcus MecA is stable in detection result, high in accuracy rate, high in sensitivity, low in cost and short in time consumptionand suitable for clinical promotion. The detection method is suitable for clinically and quickly detecting methicillin resistant staphylococcus and screening staphylococcus at pharynx nasalis, is quick and accurate, is simple in equipment, is low in requirements for operators, and has potential in bedside detection and application in hospitals at different levels.

The present invention provides improved tests for the detection of methicillin-resistant Staphylococcus aureus bearing a variant mecA gene. The tests are particularly useful for eliminating certain false negative results due to the presence of this variant in MRSA in patient samples.

The invention discloses a primer and a probe composition and a kit. The primer and probe composition are used for detecting coagulase negative staphylococcus, Staphylococcus aureus and methicillin resistance genes; a coagulase negative staphylococcus detection target is a sodA gene, the staphylococcus aureus detection target is a femA gene, and the methicillin resistance gene is a mecA gene; The primer and probe composition include a forward primer, a reverse primer, and a probe for respectively detecting the sodA gene, the femA gene, and the mecA gene, which are shown as SEQ ID NO. 1 to SEQ ID NO. 9. The primer and a probe composition use real-time fluorescent quantitative PCR to detect the coagulase negative staphylococcus, staphylococcus aureus and methicillin resistance genes. The detection cycle is greatly shortened, the efficiency and the detection flux are improved, the detection requirement is catered for, and the specificity, sensitivity and accuracy are high, at the same time, no detection process is generated after amplification in the experiment, and the pollution problem in the experiment can be effectively solved.

The invention belongs to the field of clinical examination and diagnosis, and relates to a method and kit for rapidly detecting the drug resistance and virulence of staphylococcus aureus. The kit comprises three pairs of multiple PCR amplification primers, 2*PCR premix system, and sterilized double distilled water, wherein the multiple PCR amplification primers are spa gene upstream/downstream primers, mecA gene amplification upstream/downstream primers, and pvl gene amplification upstream/downstream primers. The kit comprises three pairs of PCR primers and thus can simultaneously amplify the spa, mecA, and pvl genes of staphylococcus aureus. The kit can be used to detect staphylococcus aureus, which is resistant to methicillin, in a clinical sample, can detect staphylococcus aureus with high virulence at the same time, and provides references for the clinical treatment on staphylococcus aureus infection. The invention establishes a method for rapidly detecting drug resistance and virulence of staphylococcus aureus, and the method is simple, rapid, and cheap and has an important meaning for the clinical treatment on staphylococcus aureus infection.

The invention belongs to the field of pharmacy, and relates to a benzene polyene sesquiterpene derivative and an application thereof in preparing an anti-resistant-bacteria medicine and preparation, and especially in preparing medicines and preparations used for inhibiting drug-resistant staphylococcus aureus. The benzene polyene sesquiterpene derivative provided by the invention comprises compounds 1 and 2, and is obtained by separation from a traditional Chinese medicine material asafoetida. As a result of test, the derivative has pharmacological activity against drug-resistant staphylococcus aureus. The compound 2 has a minimal inhibitory concentration (MIC) of 2mug/L against tetracycline-resistant staphylococcus aureus strain XU212 and methicillin-resistant staphylococcus aureus strain EMRSA-15, such that the activity is substantially better than control drugs tetracycline and oxacillin. For bacterium drug resistance, the compounds 1 and 2 show significant external pump inhibition effect against resistant staphylococcus aureus RN4220 with MsrA macrolide external pump protein and methicillin-resistant staphylococcus aureus EMRSA-16 with mecA gene. The derivative can be further used for preparing medicine preparations used for resisting external-pump drug-resistant staphylococcus aureus.

The invention relates to the technical field of biomedicine, and particularly relates to a system for simultaneously detecting methicillin-susceptible and methicillin-resistant staphylococcus aureus and a using method thereof. The system comprises an m-LAMP unit and a detection unit; the m-LAMP unit is used for performing loop-mediated isothermal amplification on femA gene and mecA gene simultaneously in the same reaction system; the m-LAMP unit comprises a first primer combination for performing loop-mediated isothermal amplification on the femA gene and a second primer combination for performing loop-mediated isothermal amplification on the mecA gene; and the detection unit is used for detecting femA gene and mecA gene products after loop-mediated isothermal amplification. The system canrealize simultaneous detection of MSSA and MRSA, and can be applied to practice operations of mechanisms such as hospitals and the like for detection of MSSA and MRSA, and effective reference is provided for formulating a timely and accurate treatment scheme.

The invention discloses primer sets for detecting an mecA gene of methicillin-resistant staphylococcus aureus on a severely-infected patient, and further relates to a method for detecting the methicillin-resistant staphylococcus aureus through the primer sets and a quick quantitative detection kit for detecting the methicillin-resistant staphylococcus aureus through the primer sets. According to the primer sets, the method and the kit, the best primer sequence and the best detection method for measuring the methicillin-resistant staphylococcus aureus with the mecA gene are determined, and a quick and convenient LAMP high-throughput detection method for laboratories is built.

The invention discloses an ultra-sensitive electrochemical LCR sensor for detecting methicillin-resistant staphylococcus aureus in synovial fluid. The ultra-sensitive electrochemical LCR sensor comprises the following steps of (1) designing two specific double-stranded short probes (S-dsDNA) according to conservative series of a MecA gene; (2) in an LCR reaction system, taking the MecA gene as a template, connecting two short series of probes through DNA ligase to form long double-stranded DNA (L-dsDNA), and then carrying out cyclic amplification by using an L-dsDNA template to form a large amount of L-dsDNA, and (3) fixing the formed L-dsDNA with sulfydryl and biotin modification on a BSA modified gold electrode through a gold-sulfur bond, dropwise adding avidin-PolyHRP to be combined with biotin on the L-dsDNA, finally putting the electrode into a base solution containing TMB and H2O2, and enabling H2O2 to oxidize TMB to generate an electrochemical signal under the catalysis of HRP. The sensor has the advantages of economy, quickness, high sensitivity, specificity and the like, can be used for detecting a single base mutation series, and realizes the detection of the MRSA in the synovial fluid of a patient infected around the joint prosthesis.

A composition for diagnosing sepsis, and a method and a kit therefor. The kit includes a primer composition containing specific individual primers for amplifying: a segment of a 16s rRNA gene; a segment of an ITS gene; a segment of a Nuc gene; a segment of a MecA gene; segments of a VanA and a VanB; a segment of an invA gene; a segment of an ipaH gene; and a segment of a Cps gene. The kit also includes a probe composition containing specific individual probes for detecting: gram-negative bacteria; gram-positive bacteria; a Nuc gene; a MecA gene for checking whether or not MRSA has antibiotic resistance; a VanA and a VanB, for checking whether or not VRE have antibiotic resistance; and a gene for identifying a fungus.

The invention relates to the technical field of biological detection, in particular to primers, probes and a kit for multiplex PCR detection of seven drug-resistant genes. The kit disclosed by the invention can be used for carrying out joint detection on carbapenem enzyme genes KPC, NDM, IMP, VIM and OXA48 genes in carbapenem drug-resistant enterobacteriaceae bacteria, methicillin-resistant staphylococcus mecA genes and vanA genes in vancomycin-resistant multi-drug-resistant enterococcus in a rapid, accurate and ultrahigh-sensitivity manner; the change of the specific drug-resistant gene in the body of a blood flow infection patient can be rapidly monitored in real time, the condition of the patient is evaluated in time, and auxiliary treatment suggestions are provided for clinicians. The kit disclosed by the invention can also be used for active screening of drug-resistant genes of seven bacteria in CRE, MRSA and VRE of critically ill patients and immunosuppressive people without symptom colonization, and has extremely high application value in the field of scientific research and clinical application.

The invention discloses an application of 24-membered macrocyclic Schiff base in preparation of a drug for treating methicillin-resistant staphylococcus aureus infection. The 24-membered macrocyclic Schiff base is found to form a G-quadruplex structure with the core sequence of the MecA gene in the methicillin-resistant staphylococcus aureus through methods such as circular dichroism spectrum andWestern-Blot, so that the expression of the corresponding penicillin-binding protein PBP2a is inhibited. Pharmacological sensitivity experiments prove that the 24-membered macrocyclic Schiff base canreduce the minimum inhibitory concentration of methicillin-resistant staphylococcus aureus on oxacillin. When the use amount of the 24-membered macrocyclic Schiff base is 5 [mu] M, the minimum inhibitory concentration of methicillin-resistant staphylococcus aureus can be reduced to 0.5 [mu] g/mL to the minimum, and the effect of treating methicillin-resistant staphylococcus aureus infection by using beta-lactam drugs is achieved.