Mycobacterium Mutations Associated with Clarithromycin Resistance
|
By LabMedica International staff writers Posted on 09 Jan 2018 |
|
Image: The Sensititre OptiRead automated fluorometric plate reading system for use with Sensititre microtiter plates in antibiotic susceptibility tests (Photo courtesy of Thermo Fisher Scientific).
Mycobacterium abscessus is a leading cause of respiratory diseases and soft tissue infections and it is naturally resistant to many antibiotics in vitro, which complicates clinical treatment and leads to unsatisfactory results.
Macrolides, such as clarithromycin and azithromycin, were regarded as the foundation for the therapeutic regimen for M. abscessus complex infections. However, clarithromycin resistance is increasing dramatically among M. abscessus complex.
Microbiologists at Tongji University School of Medicine (Shanghai, China) collected specimens from the affiliated Shanghai Pulmonary Hospital and a collection of 139 M. abscessus subsp. abscessus and 36 M. abscessus subsp. massiliense clinical isolates was used to explore the utility of the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique for the rapid identification of 23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(41) (erm(41) and the 23S ribosomal RNA (rrl) gene mutations.
The team designed four primer sets to scan for mutations in the erm(41) and rrl genes using DGGE. PCR reactions and electrophoresis, as well as band retrieval and gel analysis, were performed according to previously described protocols. A Sensititre RAPMYCO panel was used to test the susceptibility of various antibiotics including clarithromycin against the 175 M. abscessus complex isolates.
The authors found a combination of 16 different DGGE patterns were observed for erm(41) gene, including 16 in M. abscessus subsp. abscessus and one in M. abscessus subsp. massiliense. Six DGGE patterns were obtained for rrl gene. Mutations in the erm(41) and rrl detected by DGGE were 100% identical to mutations detected by DNA sequencing. Among the 139 M. abscessus subsp. abscessus isolates, 36 isolates exhibited clarithromycin resistance on day 5. The other 103 were sensitive to clarithromycin on day 5, and 83 of these exhibited inducible resistance after 14 days incubation. Of the 36 M. abscessus subsp. massiliense isolates, 27 were sensitive and eight were resistant to clarithromycin on both days 5 and 14. Notably, one of the isolates was sensitive to clarithromycin on day 5, but displayed inducible resistance on day 14.
The authors concluded that their results demonstrated that DGGE technology is very sensitive to point mutations, as well as to small insertions and deletions in DNA sequences. The accuracy was essentially 100% among targeted fragments of the erm(41) and rrl genes. The results of whole genome sequencing confirmed DGGE efficacy, all distinct nucleotides polymorphisms were detected. PCR-based DGGE is a practical technique for the rapid detection of mutations in the erm(41) and rrl genes associated with clarithromycin resistance in M. abscessus complex. The study was published in the December 2017 issue of the Journal of Microbiological Methods.
Related Links:
Tongji University School of Medicine
Macrolides, such as clarithromycin and azithromycin, were regarded as the foundation for the therapeutic regimen for M. abscessus complex infections. However, clarithromycin resistance is increasing dramatically among M. abscessus complex.
Microbiologists at Tongji University School of Medicine (Shanghai, China) collected specimens from the affiliated Shanghai Pulmonary Hospital and a collection of 139 M. abscessus subsp. abscessus and 36 M. abscessus subsp. massiliense clinical isolates was used to explore the utility of the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) technique for the rapid identification of 23S rRNA (Adenine(2058)-N(6))-methyltransferase Erm(41) (erm(41) and the 23S ribosomal RNA (rrl) gene mutations.
The team designed four primer sets to scan for mutations in the erm(41) and rrl genes using DGGE. PCR reactions and electrophoresis, as well as band retrieval and gel analysis, were performed according to previously described protocols. A Sensititre RAPMYCO panel was used to test the susceptibility of various antibiotics including clarithromycin against the 175 M. abscessus complex isolates.
The authors found a combination of 16 different DGGE patterns were observed for erm(41) gene, including 16 in M. abscessus subsp. abscessus and one in M. abscessus subsp. massiliense. Six DGGE patterns were obtained for rrl gene. Mutations in the erm(41) and rrl detected by DGGE were 100% identical to mutations detected by DNA sequencing. Among the 139 M. abscessus subsp. abscessus isolates, 36 isolates exhibited clarithromycin resistance on day 5. The other 103 were sensitive to clarithromycin on day 5, and 83 of these exhibited inducible resistance after 14 days incubation. Of the 36 M. abscessus subsp. massiliense isolates, 27 were sensitive and eight were resistant to clarithromycin on both days 5 and 14. Notably, one of the isolates was sensitive to clarithromycin on day 5, but displayed inducible resistance on day 14.
The authors concluded that their results demonstrated that DGGE technology is very sensitive to point mutations, as well as to small insertions and deletions in DNA sequences. The accuracy was essentially 100% among targeted fragments of the erm(41) and rrl genes. The results of whole genome sequencing confirmed DGGE efficacy, all distinct nucleotides polymorphisms were detected. PCR-based DGGE is a practical technique for the rapid detection of mutations in the erm(41) and rrl genes associated with clarithromycin resistance in M. abscessus complex. The study was published in the December 2017 issue of the Journal of Microbiological Methods.
Related Links:
Tongji University School of Medicine
Gold Member
Troponin T QC
Troponin T Quality Control
New
Myeloperoxidase Assay
IDK MPO ELISA
New
H.pylori Test
Humasis H.pylori Card
Latest Microbiology News
- Handheld Device Deliver Low-Cost TB Results in Less Than One Hour
- New AI-Based Method Improves Diagnosis of Drug-Resistant Infections
- Breakthrough Diagnostic Technology Identifies Bacterial Infections with Almost 100% Accuracy within Three Hours
- Innovative ID/AST System to Help Diagnose Infectious Diseases and Combat AMR
- Gastrointestinal Panel Delivers Rapid Detection of Five Common Bacterial Pathogens for Outpatient Use
- Rapid PCR Testing in ICU Improves Antibiotic Stewardship
- Unique Genetic Signature Predicts Drug Resistance in Bacteria
- Unique Barcoding System Tracks Pneumonia-Causing Bacteria as They Infect Blood Stream
- Rapid Sepsis Diagnostic Test Demonstrates Improved Patient Care and Cost Savings in Hospital Application
- Rapid Diagnostic System to Detect Neonatal Sepsis Within Hours
- Novel Test to Diagnose Bacterial Pneumonia Directly from Whole Blood
- Interferon-γ Release Assay Effective in Patients with COPD Complicated with Pulmonary Tuberculosis
- New Point of Care Tests to Help Reduce Overuse of Antibiotics
- 30-Minute Sepsis Test Differentiates Bacterial Infections, Viral Infections, and Noninfectious Disease
- CRISPR-TB Blood Test to Enable Early Disease Diagnosis and Public Screening
- Syndromic Panel Provides Fast Answers for Outpatient Diagnosis of Gastrointestinal Conditions
Channels
Clinical Chemistry
view channel
Carbon Nanotubes Help Build Highly Accurate Sensors for Continuous Health Monitoring
Current sensors can measure various health indicators, such as blood glucose levels, in the body. However, there is a need to develop more accurate and sensitive sensor materials that can detect lower... Read more
Paper-Based Device Boosts HIV Test Accuracy from Dried Blood Samples
In regions where access to clinics for routine blood tests presents financial and logistical obstacles, HIV patients are increasingly able to collect and send a drop of blood using paper-based devices... Read more
Molecular Diagnostics
view channel
RNA-Based Blood Test Detects Preeclampsia Risk Months Before Symptoms
Preeclampsia remains a major cause of maternal morbidity and mortality, as well as preterm births. Despite current guidelines that aim to identify pregnant women at increased risk of preeclampsia using... Read more
First Of Its Kind Test Uses microRNAs to Predict Toxicity from Cancer Therapy
Many men with early-stage prostate cancer receive stereotactic body radiotherapy (SBRT), a highly precise form of radiation treatment that is completed in just five sessions. Compared to traditional radiation,... Read more
Novel Cell-Based Assay Provides Sensitive and Specific Autoantibody Detection in Demyelination
Anti-myelin-associated glycoprotein (MAG) antibodies serve as markers for an autoimmune demyelinating disorder that affects the peripheral nervous system, leading to sensory impairment. Anti-MAG-IgM antibodies... Read more
Hematology
view channel
New Scoring System Predicts Risk of Developing Cancer from Common Blood Disorder
Clonal cytopenia of undetermined significance (CCUS) is a blood disorder commonly found in older adults, characterized by mutations in blood cells and a low blood count, but without any obvious cause or... Read more
Non-Invasive Prenatal Test for Fetal RhD Status Demonstrates 100% Accuracy
In the United States, approximately 15% of pregnant individuals are RhD-negative. However, in about 40% of these cases, the fetus is also RhD-negative, making the administration of RhoGAM unnecessary.... Read more
Immunology
view channel
Stem Cell Test Predicts Treatment Outcome for Patients with Platinum-Resistant Ovarian Cancer
Epithelial ovarian cancer frequently responds to chemotherapy initially, but eventually, the tumor develops resistance to the therapy, leading to regrowth. This resistance is partially due to the activation... Read more
Machine Learning-Enabled Blood Test Predicts Immunotherapy Response in Lymphoma Patients
Chimeric antigen receptor (CAR) T-cell therapy has emerged as one of the most promising recent developments in the treatment of blood cancers. However, over half of non-Hodgkin lymphoma (NHL) patients... Read more
Pathology
view channel
Advanced Imaging Reveals Mechanisms Causing Autoimmune Disease
Myasthenia gravis, an autoimmune disease, leads to muscle weakness that can affect a range of muscles, including those needed for basic actions like blinking, smiling, or moving. Researchers have long... Read more
AI Model Effectively Predicts Patient Outcomes in Common Lung Cancer Type
Lung adenocarcinoma, the most common form of non-small cell lung cancer (NSCLC), typically adopts one of six distinct growth patterns, often combining multiple patterns within a single tumor.... Read more
Technology
view channel
Pain-On-A-Chip Microfluidic Device Determines Types of Chronic Pain from Blood Samples
Chronic pain is a widespread condition that remains difficult to manage, and existing clinical methods for its treatment rely largely on self-reporting, which can be subjective and especially problematic... Read more
Innovative, Label-Free Ratiometric Fluorosensor Enables More Sensitive Viral RNA Detection
Viruses present a major global health risk, as demonstrated by recent pandemics, making early detection and identification essential for preventing new outbreaks. While traditional detection methods are... Read more
Industry
view channel
Cepheid and Oxford Nanopore Technologies Partner on Advancing Automated Sequencing-Based Solutions
Cepheid (Sunnyvale, CA, USA), a leading molecular diagnostics company, and Oxford Nanopore Technologies (Oxford, UK), the company behind a new generation of sequencing-based molecular analysis technologies,... Read more
Grifols and Tecan’s IBL Collaborate on Advanced Biomarker Panels
Grifols (Barcelona, Spain), one of the world’s leading producers of plasma-derived medicines and innovative diagnostic solutions, is expanding its offer in clinical diagnostics through a strategic partnership... Read more
