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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



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



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



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.



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.



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 ([email protected])

CambodiaRodolphe Mérieux Laboratory, University of Health Sciences, Phnom Penh

Monidarin Chou ([email protected])

Cameroon Biotechnology centre of Yaoundé I University

Wilfred Mbacham ([email protected])

Canada Luminex Molecular Diagnostics, Inc, Toronto

Richard Janesczko ([email protected])

China Christophe Mérieux Laboratory, Chinese Academy of Medical Sciences (CAMS) and Institute of

Pathogen Biology (IPB), Beijing

Jianwei Wang ([email protected])

France Emerging Pathogens Laboratory, Fondation Mérieux, Lyon

Glaucia Baccalà ([email protected])

Haiti Rodolphe Mérieux Laboratory, GHESKIO Centres, Port au Prince

Jean William Pape ([email protected])



LaosInfectiology Centre Christophe Mérieux, Vientiane

Phimpha Paboriboune ([email protected])

Lebanon Rodolphe Mérieux Laboratory of Saint Joseph University, Beirut

Dolla Sarkis ([email protected])

Luxembourg Fast-Track Diagnostics, Junglinster

Bill Carman ([email protected])

Madagascar Infectiology Centre Charles Mérieux of Antananarivo University

Arsène Ratsimbasoa ([email protected])

Mali Infectiology Centre Charles Mérieux of Bamako

Souleymane Diallo ([email protected])

MongoliaMongolian Academy of Medical Sciences, Ulaanbaatar

Pagbajab Nymadawa ([email protected])

ParaguayInstituto de Investigaciones en Ciencias de la Salud, National University of Asunción

Graciela Russomando ([email protected])

GABRIEL Network Manager

Florence Pradel

[email protected]

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