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Lab-on-Chip-Based Platform for Fast Molecular Diagnosis of Multidrug- Resistant Tuberculosis
PRESENTED BY :DR SEEMA ALEEM
MODERATOR: DR.VARSHA.A. SINGH
AUTHORS :Andrea M. Cabibbe, Paolo Miotto, Raquel Moure, Fernando Alcaide, Silke Feuerriegel, Gianni Pozzi, Vladislav Nikolayevskyy, Francis Drobniewski, Stefan Niemann, Klaus Reither, Daniela M. Cirillo
PLACE: Sambya Hospital, Kampala, UgandaJOURNAL: Journal of Clinical MicrobiologyVOLUME: December 2015 Volume 53 Number 12, page no:3876-3880introduction MDR-TBlong and expensive treatment/ poor clinical outcomeWHO -molecular diagnostics for fast diagnosis Mycobacterium tuberculosis complex (MTBC) Genotypic diversity and geographical distribution Silent mutations The limited versatility are some of the restraints undermining the effectiveness of the current tools on a global scale.Molecular lab-on-chip-based VerePLEX Biosystem for detection of MDR-TBDiagnostic accuracy > 97.8% Compared to sequencing and MTBDRplus assay Rifampicin and isoniazid resistance- smear-positive specimens. The speed, user-friendly interface, and versatility .
principalAim and objectivesTo evaluate the different mycobacterial species with the targeted probes.To evaluate different patterns of mutations in rpoB, katG, and inhA genesprincipalMATERIAL AND METHOD Genes used: rpoB katG inhA
RESULTS rpoB - 99.00%, katG - 95.80% inhA - 95.50% targets, respectively. 100% sensitivity and specificity - Lab on chip, resistance to RIF
CONCLUSION
high specificity and sensitivity of the semi automated VerePLEX Biosystem for the MDR-TB targets, thus suggesting an usefulness of the platform for fast and simple diagnosis of MDR cases in centralized laboratories.principalVereplex biosystemVerePLEX - Versatile multiplexing platform Qualitative analysis of nucleic acids Lab-on-chip (LoC) device: developed by STMicroelectronics (Geneva,Switzerland)marketed by Veredus Laboratories (Republic of Singapore) as the VerePLEX Biosystem, for the diagnosis of MDR-TB and detection of common non-tuberculous mycobacteria (NTM).principalprincipal It integrates two powerful molecular biological applications: Polymerase Chain Reaction (PCR) for DNA amplificationmicroarray technology for multiplexed detection, onto a Lab-on-Chip platform.
PCR
MicroarrayLabelled nucleic acids(target) in a solution hybridize
Complementary sequences (probe) on solid sustrate ie.,chip
Generate signals
Detected by scanner & quantified by image analyser software
22VerePELEX Human sample bacterial DNA extraction
PCR of extracted bacterial DNA using vere lab on chip and specific kits
Hybridization of PCR product on the microarray
Washing of the microarray
Analysis of microarray results
COMPONENTSTEMPERATURE CONTROLCompletely independent , thermally drives
five different tests simultaneously accommodate the needs of both large diagnostic settings and decentralized settings at the point-of-needTemperature control - extremely accurate -very high heating / cooling ramp rates -PCR protocols - 50-75% shorter times than conventional thermal cyclers.OPTICAL READERCompact
Easy-to-use operating mode
Detects and analyzes the microarray fluorescence in between 670 and730 nm in a few seconds; automatically selected minimally-trained personnel
point-of-need applications.softwareTo support research on genomics and implementation of products and services based on microarrays. Sophisticated and advanced algorithms are used for capture, processing and interpretation of genomic data obtained through microarray technology.Instrument control and data analysis in one package.Automated startup and easy connection with the instruments.User friendly.
components of VerePLEX Software Package:E@syControl controls the Temperature Control System. It allows the user to:
Calibrate the thermometric chain.
Select the temperature program before each run
Monitor the temperature profile during run
Obtain log data from the instrumentation when needed
Control up to 5 Temperature Control Systems simultaneously
Lock each module of the Temperature Control System during operation
E@syCheck drives the Optical Reader.It allows the user to:Acquire and analyze microarray image
Save the data
Generate report
Review and export raw data
Reanalyze the microarray image
Within the software, each application is bundled as a BioApplication.
E@syManagerallows advance user to:
View and manage the user account information
Manage Bio Applications
Show currently using the TCS
View and export the TCS chips data traceability
View and export the Optical Reader chips data traceability
Create new views for data visualization
principalADVANTAGES Enable Better Decisions: Rapid answers
Ease-of-Use: Miniaturization, integration, and automation to allow minimally trained personnel
Multiplexing: The ability to assess many analytes in a single test.
Versatility: Compatible with existing assay technologies.
Diverse Market Segments: Ideal for large number of molecular biological testing sites.VereFLU
PROBES FOR:Influenza A type (type probes for H1, H3, H5, H7, H9)Influenza A subtypes (specific probes for subtypes: H1N1, A/H1N1-2009, H3N2, H5N1, H9N2)newly emergent Influenza AAdditional identification of H5N1 using specific probesInfluenza B
SAMPLE TYPES: Blood Respiratory swabs : nasopharyngeal, throat, nasal aspirates
VereTropVereTrop D and VereTrop R: allow for rapid detection, differentiation identification of major bacterial and viral tropical diseases, respectively.
VereTHREAT
VereThreat is a nucleic acid-based, Lab-On-Chip (LOC) device which combines multiplex PCR and microarray hybridization to detect, differentiate and identify :Anthrax, Small Pox, Plague Tularemia.
Detects four selected Category A biological agents Multiple probes (with duplicates) for:Bacillus anthracisFrancisella tularensisYersinia pestisVariola virus
Sample Types: Air, Water, Soil Food
VereMERSDetection of NS3 and S target genes with multiple probes to positively identify MERS-CoV
Sample Types: Serum Respiratory swabs : Nasopharyngeal Oropharyngeal Throat Nasal aspiration
