Lab-on-Chip-Based Platform for Fast

Molecular Diagnosis of MultidrugResistant 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, Uganda

JOURNAL: Journal of
VOLUME: December 2015

Clinical Microbiology

Volume 53 Number 12, page no:3876-3880

INTRODUCTION

 MDR-TBlong and expensive treatment/ poor clinical

outcome
WHO -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-TB
Diagnostic accuracy > 97.8%
Compared to sequencing and MTBDRplus assay
Rifampicin and isoniazid resistance- smear-positive
specimens.
The speed, user-friendly interface, and versatility .

AIM
AND
OBJECTIVES
principal

 To evaluate the different mycobacterial species with the

targeted probes.
To evaluate different patterns of mutations in rpoB, katG,
and inhA genes

MATERIAL AND
METHOD
principal

Culture negative

METHODS
VerePLEX
COMPARED
Biosystem

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.

VerePLEX
BIOSYSTEM
principal

VerePLEX - 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).

PRINCIPAL
principal

It integrates two powerful molecular biological

applications:
Polymerase Chain Reaction (PCR) for DNA
amplification
microarray

technology

detection,
onto a Lab-on-Chip platform.

for

multiplexed

PCR

Microarray
Labelled 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

22

VerePELEX
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

COMPONENTS
Optical reader

TEMPERATURE CONTROL
Completely independent , thermally drives

five different tests simultaneously

accommodate the needs of both large diagnostic settings

and decentralized settings at the point-of-need
Temperature control - extremely accurate
-very high heating / cooling ramp
rates
-PCR protocols - 50-75% shorter times than
conventional thermal cyclers.

OPTICAL READER
Compact
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.

software
To 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@syManager allows 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

ADVANTAGES
principal

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 A

Additional identification of H5N1 using specific probes

Influenza B
SAMPLE TYPES:
Blood
Respiratory swabs :

nasopharyngeal,
throat,
nasal aspirates

VereTrop

VereTrop 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 anthracis
Francisella tularensis
Yersinia pestis
Variola virus
Sample Types: Air,
Water,
Soil
Food

VereMERS

Detection of NS3 and S target genes with multiple probes to positively

identify MERS-CoV
Sample Types:

Serum
Respiratory swabs : Nasopharyngeal
Oropharyngeal
Throat
Nasal aspiration