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www.gabriel-network.org Issue 8
Global Approach for Biological Research on Infectious Epidemics in Low income countries 1/7
MEMBERS NEWSKeeping you updated!
Contents
The Rodolphe Mérieux
Laboratory project in
Bangladesh
Beyond species, TB
genotyping
Diagnostic biomarkers
for fevers in children
living in a malaria-
endemic region
The 4th HIV Workshop
in Luang Prabang, Laos:
Therapeutic failure
and early diagnosis for
children
A Pilot study: Prevalence
of selected waterborne
pathogens in various
water sources in Phnom
Penh, Cambodia
A team from Fondation
Mérieux visits the
GHESKIO centres in Haïti
GABRIEL Network
members and contacts
The Rodolphe Mérieux Laboratory project in
Bangladesh Following two preparatory missions conducted by
Fondation Mérieux on the recommendations of the
Wellcome Trust representative in Thailand, the Fondation
Mérieux and the Ministry of Health have jointly signed
an agreement for the creation of a Rodolphe Mérieux
laboratory in Chittagong, Bangladesh’s second largest
city after Dhaka.
The laboratory will be located within the Bangladesh Institute for
Tropical Institute Diseases (BITID) currently being built in Chittagong.
The BITID is a research institute and a health centre mainly devoted
to infectious diseases: enteric diseases, (especially typhoid)
tuberculosis, malaria (prevalent in the forested areas close to the
city), respiratory infections, parasitic diseases, and rabies.
The seven-story building will be able to accommodate up to 200
researchers and health personnel. The first phase of construction,
a four-story building, is to be operational in late 2011 or early 2012.
A review team from Fondation Mérieux has assessed the progress
of the construction of the RML, and a study of potential scientific
collaborative initiatives was carried out in January 2011. The 4th floor
laboratory will initially consist of a BSL-2 and biological laboratories
that are to include molecular biology. A BSL-3 is planned at a later
stage. Development plans have been submitted to the directors of
the BITID (Pr. Abul Faiz and Dr. M. Amir Hossain) and discussions
are currently underway. The Scientific Department of the Fondation
Mérieux will have the responsibility of advising and supervising future
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Global Approach for Biological Research on Infectious Epidemics in Low income countries 2/7
work projects. The topics, “respiratory infections” and “typhoid”, have been selected to launch the start
of cooperative scientific initiatives within the scope of the GABRIEL Network. A contract governing the
various activities to be conducted is expected to be signed soon with the Bangladesh Ministry of Health.
Guy Vernet, Fondation Mérieux, France
Beyond species, TB genotypingThe strains of M. tuberculosis found in the world’s population exhibit varying clinical features in terms of
their mode of transmission, virulence, and the degree to which they are likely to develop drug resistance.
The origin of these differences can be traced by studying their bacterial genome. Molecular markers,
such as locus DR, MIRU, IS6110, are presently used to group strains into clades. The Hain-Lifescience kit is
used to identify species and test their resistance to drugs. However, going one step further, the Emerging
Pathogens Laboratory has developed a molecular assay to actually define the genotype of strains by
spoligotype analysis using the locus DR marker. It is expected that this molecular tool, from a single test
tube, will be able to:
• confirm the presence of a species of the MTB complex species,
• identify the species within the complex,
• examine mutations that confer resistance to Rifampicin and Isoniazid, and
• define the spoligotype.
This work is being carried out in collaboration with Prof. C. Sola’s team at the Université Paris-Sud, one PhD
student of which is being funded by the Fondation Mérieux. Gene amplification is carried out by PCR, and
detection is performed using Luminex’s MagPix system: hybridization probes to target DNA sequences
are bound to magnetic and fluorescent beads. This system can analyze up to 50 single probes.
The typing of strains prevalent in the GABRIEL Network member countries should lead to the identification
of genotypes and to the association of each with specific clinical characteristics, such as drug resistance.
In addition, the most widespread genotypes in the population will be identified and thus singled out as the
cause of epidemics.
Jean-Luc Berland, Emerging Pathogens Laboratory, Fondation Mérieux, France
Diagnostic biomarkers for fevers in children living in a malaria-endemic regionA study on diagnostic biomarkers for fevers in
children was launched in May 2011 as part of
the applied research activities of the Fondation
Mérieux, coordinated by the Emerging Pathogens
Laboratory, the Institut Mérieux, Caprion,
the Charles Mérieux Centre of Infectiology in
Madagascar and the NIAID / NIH.
This prospective study for the detection of
biomarkers and the discrimination of various febrile
illnesses due to malaria/ALRI is being carried out,
with the collaboration of the local hospital, on a
target population of children under 5 years old
living in the malaria-endemic region of Ampasimanjeva, Madagascar. The study applies a protocol that
has received central and site IRB/IRE approval and is carried out under Good Clinical Practices (GCP) and
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Global Approach for Biological Research on Infectious Epidemics in Low income countries 3/7
SOP compliance. The goal of this study is to identify, by mass spectrometry, protein biomarkers that are
specifically induced by causative pathogens of fevers (bacteria, viruses and/or atypical bacteria, parasites)
in a malaria-endemic region. This study should provide a greater understanding of causative biomarkers
of these diseases, so that more accurate diagnostic procedures to distinguish the overlapping of febrile
illnesses can be developed.
Glaucia Baccalà, Emerging Pathogens Laboratory (EPL), Fondation Mérieux, France
The 4th HIV Workshop in Luang Prabang, Laos: Therapeutic failure and early diagnosis for childrenThe 4th HIV workshop that was held in Luang Prabang
in the Laos, from 4-6 May 2011, focused on the early
diagnosis of HIV infection in children less than 18
months of age. This workshop follows an initiative
started in November 2009 from clinicians, who
wished to have the opportunity to meet biannually to
talk about issues of common interest.
Various regional and international organizations
participated in the organization: Centre for HIV/AIDS
and STI (CHAS), Christophe Mérieux Centre of Laos
(CCML), the Global Fund, ESTHER (Laos), the Institut
Pasteur of Cambodia, and Fondation Mérieux.
The workshop began with presentations by the organizers and health professionals involved in HIV /
AIDS, followed by a review of the work that has been carried out by the CCML on HIV viral load and drug
resistance.
During the workshop, the representatives from France, Cambodia, and Vietnam spoke about their
experiences in the use of tests with dry blood spot samples obtained from heel pricks. From discussions
that followed on the approach for early HIV diagnosis, participants came up with measures that will provide
for the care of children born to HIV-positive mothers. Of major importance are the steps that must be taken
to prevent HIV transmission from mother to child, shorten the delay between diagnosis and treatment,
and define the kind of first-line treatment that should be administered after ingestion of Nevirapine. Also
discussed was the matter regarding the time span between each viral load count performed for newborn
infants, given that HIV treatment in the past has largely failed due to pre and post resistance to ARV.
Clinicians and pediatricians from seven HIV/AIDS treatment centres in Laos participated at this workshop,
as well as supervisors from CHAS. Among the principal moderators, Dr. Phimpha Paboriboune, Scientific
Director of the CCML, took part in the discussions and helped organize this event. Several foreign participants
contributed to enliven the debates and shared their experiences: Prof. Diane Descamps (Bichat Hospital,
Paris, France), Dr. Christophe Longuet (Fondation Mérieux, Lyon, France), Dr. Eric Nerrienet (IPC, Phnom
Penh, Cambodia), and Dr. Marcelo Fernandez (HAIVN, Hanoi, Vietnam).
All the participants expressed their interest in attending future workshops. The next workshop on the
theme of HIV-TB and HIV-Hepatitis co-infections will be held late 2011 or early 2012.
Fondation Mérieux is the partner of choice for the organization of these workshops and has contributed to
the introduction and setting up of viral load counts in Laos, therefore playing an important role in the fight
against HIV in Laos.
Vibol IEM, Christophe Mérieux Centre of Laos
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Global Approach for Biological Research on Infectious Epidemics in Low income countries 4/7
A Pilot study: Prevalence of selected waterborne pathogens in various water sources in Phnom Penh, Cambodia
The World Health Organisation has reported that
among neonates and children under five years old,
diarrhea remained a global problem accounting for
14% mortalities in 2008. For example, in Southeast
Asia alone, diarrhea diseases are responsible for
about 12% of all post neonate deaths. A vast number
of pathogens, ranging from viruses to bacteria to
parasites (e.g. protozoan and helminths), have
been detected in different types of water used for
human activities, and these generally gave rise to
enteric diseases.
In order to investigate the burden of diarrhea
diseases in Cambodian children, the Rodolphe Mérieux Laboratory of Phnom Penh started a study in 2009
on the presence of common water-borne pathogens in water used for different types of daily activities.
In this pilot study, water from 10 selected sites within 20 km of Phnom Penh were sampled. Seven of these
sites were along the Mekong and Tonlé Sap rivers, and three other sites studied samples from a pond,
well and reservoir. Twenty litres of water were sampled from each site during the dry and rainy seasons.
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Global Approach for Biological Research on Infectious Epidemics in Low income countries 5/7
Each sample collected was first concentrated using the tangential flow filtration followed by serial filtration
with decreasing membrane size from 1 µm to 0.1 µm. Nucleic acids were extracted from each fraction
and subjected to molecular analysis by end-point Polymerase Chain Reaction (PCR). It was observed that
the Cambodians used untreated water directly from natural sources, rivers, ponds, and underground.
The presence of Leptospira spp. was detected from all sites and also other pathogens including E.
coli, Salmonella typhi, Shigella flexneri, Vibrio cholerae, Vibrio mimicus and Vibrio parahaemolyticus. This
preliminary data is currently being confirmed by other molecular-based analysis and bacterial isolation
techniques.
This data suggests that enteric pathogens are generally present in untreated water in Cambodia and
explains the prevalence of diarrhea diseases in Cambodian children. This also explains the increase in
enteric epidemics as reported in the recent message from ProMed. The data further suggests that all
forms of untreated water should be processed before being used for daily activities.
Monidarin Chou, Rodolphe Mérieux Laboratory of Cambodia and Boon-Huan Tan, National University
of Singapore
A team from Fondation Mérieux visits the GHESKIO Centres in Haïti
In early July 2011 a team from Fondation Mérieux
paid a one week visit to Haiti. The team was
composed of Valentina Picot (Scientific Advisor),
Sandra Dollet (Laboratory Technician) and Louis
Delorme (Project Manager). The team traveled to
the GHESKIO Centres in Port-au-Prince to conduct
three activities. A Laboratory audit was carried
out as part of the pneumonia study set up by
Fondation Mérieux. Molecular biology training for
4 laboratory technicians was implemented at the
Institute of Infectious Diseases and Reproductive
Health (IMIS). A training session for the administration and supervision of the ReST-ACMI project took place
for the local team in charge of this project.
AuditThe pneumonia study audit carried out in Haiti aimed at harmonizing all the research in order to comply
with standard operating procedures . The audit took place at the GHESKIO site for clinical part and at the
IMIS for the laboratory part. The sites met all the conditions necessary to begin the study and only requires
the approval from Ethics Committee of Cornell University which is expected for next month. The team is
highly motivated to start the study.
ReST-ACMI ProjectThe supervision of the ReST-ACMI project took place at the GHESKIO centres for four days. Meetings
and work sessions on administrative and financial points were organized with the local team and also on
the progress and planning of activities.This was also the opportunity to organize the international steering
committee of the project. This meeting helped to better understand the real needs of the GHESKIO centres
and adapt as much as possible the project to the local constraints.
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Global Approach for Biological Research on Infectious Epidemics in Low income countries 6/7
TrainingFour people of the IMIS molecular biology team
were trained in the techniques used in the
multi-centric study project on pneumonia. The
programme consisted of the extraction of nucleic
acid from nasopharyngeal samples, the real-time
PCR method for the detection of the respiratory
pathogens using the Fast Track respiratory 21 Plus
kit and the analysis of the results. The participants
were very interested and motivated during all the
training.
Despite a very difficult environment, the GHESKIO team procudes a high-quality work. Thanks to their
motivation and availability, all the objectives of the mission were achieved.
Sandra Dollet, Valentina Picot and Louis Delorme, Fondation Mérieux, France
GABRIEL Network members and contacts
BrazilRespiratory virus Laboratory of Oswaldo Cruz Institute, Fondation Fiocruz, Rio de Janeiro
Marilda Siquiera (mmsiq@ioc.fiocruz.br)
CambodiaRodolphe Mérieux Laboratory, University of Health Sciences, Phnom Penh
Monidarin Chou (cmonidarin@rmerieux-lab-cambodge.org)
Cameroon Biotechnology centre of Yaoundé I University
Wilfred Mbacham (wfmbacham@yahoo.com)
Canada Luminex Molecular Diagnostics, Inc, Toronto
Richard Janesczko (richardj@luminexcorp.com)
China Christophe Mérieux Laboratory, Chinese Academy of Medical Sciences (CAMS) and Institute of
Pathogen Biology (IPB), Beijing
Jianwei Wang (wangjw28@163.com)
France Emerging Pathogens Laboratory, Fondation Mérieux, Lyon
Glaucia Baccalà (glaucia.baccala@fondation-merieux.org)
Haiti Rodolphe Mérieux Laboratory, GHESKIO Centres, Port au Prince
Jean William Pape (jwpape@gheskio.org)
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Global Approach for Biological Research on Infectious Epidemics in Low income countries 7/7
LaosInfectiology Centre Christophe Mérieux, Vientiane
Phimpha Paboriboune (ppaboriboune@rmerieux-lab-laos.org)
Lebanon Rodolphe Mérieux Laboratory of Saint Joseph University, Beirut
Dolla Sarkis (dolla.sarkis@usj.edu.lb)
Luxembourg Fast-Track Diagnostics, Junglinster
Bill Carman (w.carman@vir.gla.ac.uk)
Madagascar Infectiology Centre Charles Mérieux of Antananarivo University
Arsène Ratsimbasoa (arsene.ratsimbasoa@laposte.net)
Mali Infectiology Centre Charles Mérieux of Bamako
Souleymane Diallo (souleymane.diallo@fondationmerieux-mali.org)
MongoliaMongolian Academy of Medical Sciences, Ulaanbaatar
Pagbajab Nymadawa (nymadawa@gmail.com)
ParaguayInstituto de Investigaciones en Ciencias de la Salud, National University of Asunción
Graciela Russomando (grusso@rieder.net.py)
GABRIEL Network Manager
Florence Pradel
florence.pradel@fondation-merieux.org
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