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Antibiotic Resistance Genes (ARGs) Analysis Solution
SYBR Green I-based qPCR Assay for ARGs

SYBR Green I-based qPCR Assay for ARGs

Overview

Antibiotic resistance is a growing problem in the world today. The overuse of antibiotics has led to the development of antibiotic-resistant bacteria, which can be difficult to treat. Multi-drug resistant E. coli, multi-drug resistant Salmonella, etc. have become the main culprits for food and human safety. The accurate and rapid detection of drug-resistance genes in microorganisms is important for guiding drug use and biosafety. CD Genomics offers comprehensive antibiotic resistance gene (ARGs)detection solutions to help combat this problem, including SYBR Green I-based qPCR assay service. SYBR Green I is a DNA dye that is simple to use and relatively low cost.

Our Advantages of SYBR Green I:

The SYBR Green I dye method does not require the use of probes and is cost-effective.
This method does not need to add a fluorescent reporter system in the primer design, which is more convenient and simpler.
SYBR Green can be used to monitor the amplification of any double-stranded DNA sequence.
No need to synthesize fluorescently labeled probes or beacons specific to the target sequence.

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Introduction to Our SYBR Green I-based qPCR Analysis Platform

SYBR Green I-based qPCR Assay for ARGs

The SYBR Green I assay is a powerful tool for detecting antibiotic resistance genes in bacteria. It is based on real-time PCR technology and uses SYBR Green I fluorescent dye to detect the presence of antibiotic resistance genes. The assay offers the possibility of simultaneous discrimination of the mutation in codon 460 and quantitative detection of the copies in mixtures. After embedding into the double-stranded DNA molecule in the PCR reaction system, an excess of SYBR fluorescent dye is added, which is specifically incorporated into the DNA double-strand and changes its conformation, absorbing 497 nm excitation light and emitting 520 nm fluorescence; whereas the dye molecule not incorporated into the DNA double-strand does not emit any fluorescence signal, thus ensuring that the increase in fluorescence signal is perfectly synchronized with the increase in the PCR product. This ensures that the increase in fluorescence signal is perfectly synchronized with the increase in the PCR product. CD Genomics offers characterization services for this assay, which can help researchers to better understand the genetic basis of antibiotic resistance.

Workflow of SYBR Green I-based qPCR Analysis of ARGs

The Flowchart of SYBR Green I-based qPCR Assay for ARGs - CD Genomics.

What We Offer

CD Genomics, as a specialist provider of antibiotic resistance gene testing services, has a strong capability to provide you with SYBR Green I-based qPCR analysis services for hundreds of antibiotic resistance genes. We have listed some of the targets that can be targeted. For more information, please contact us directly.

Gene	Classification	Mechanism
catA1, catB3, cfr, etc	(flor)/(chlor)/(am)phenicol	deactivate
cmlA1, cmx(A), floR, etc		efflux
qnr, etc		-
aac, aacA/aphD, aacC, aacC1, aacC2, aacC4, aadA, aadD, aadE, aph, aph6ia, aphA1(akakanR), spcN-01, spcN-02, str, strA, strB, etc	Aminoglycosides	deactivate
tetA, tetB, tetC, tetD, tetE, tetG, tetH, tetJ, tetK, tetL, tetPA, tet, tetV, etc	Tetracyclines	efflux

Bioinformatics Analysis

Depending on your needs, we will develop a tailor-made bioinformatics analysis solution for you.

ANALYSIS CONTENTS	DETAILS
Multiplex SYBR Green Real-Time PCR	The ability to simultaneously analyse different antibiotic resistance genes such as pbp2b, ermB and mef genes.
Relative gene expression	The relative expression of antibiotic resistance genes can be analyzed.
Absolute quantification	Absolute quantification of antibiotic resistance gene abundance.
Allelic discrimination	Alleles that can distinguish antibiotic resistance genes.
Melting curve analysis	Melting curve analysis can be used to assess qPCR amplicon length, SNP/mutation, etc.

Sample Requirements

1. DNA: 1.8 < OD260/280 < 2.0, no contamination or degradation. Please guarantee that specimens are properly secured and packaged during transport.

Sampling kits: We provide a range of microbial sampling kits for clients, including MicroCollect™ oral sample microbial collection products and MicroCollect™ stool sample collection products.

References

Fan W, Hamilton T, Webster-Sesay S, et al. Multiplex real-time SYBR Green I PCR assay for detection of tetracycline efflux genes of Gram-negative bacteria. Mol Cell Probes. 2007 Aug; 21(4): 245-56.
Souza MB, Cergole-Novella MC, Molinari DA, et al. Multiplex real-time PCR using SYBR Green: Unspecific intercalating dye to detect antimicrobial resistance genes of Streptococcus pneumoniae in cerebrospinal fluid. PLoS One. 2022 Jun 14; 17(6): e0269895.