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ប្រទេសកម្ពុជា
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醫學生物學實驗室

Since its creation in 1995, the Medical Laboratory also known as “Laboratoire de Biologie Médicale” (LBM) of the Institut Pasteur du Cambodge provides a platform for comprehensive medical and biological analyses to Cambodian and international clients

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ប្រជាជនកម្ពុជា
ENGLISH
FRANÇAIS

服務

醫學生物學實驗室

INTERNSHIP

PUBLICATIONS

Employing around 35 health professionals (Medical biologists, Medical doctors, Engineers, Technicians, Nurses, Secretaries), LBM aims at supporting health professional by offering them a variety of in vitro diagnostic tests covering all medical specialties and by permanently improving its service.

Always at the forefront of a quality service, LBM has set up its Quality Management System in order to ensure the required quality standard at pre-analytical (sample collection and transportation), analytical (testing) and post-analytical (result delivery and interpretation) level, as well as the continual improvements to ensure consistent quality results. Our main objective is to meet the expectation and satisfaction of all our clients. With our strong involvement in Quality Approach, LBM has become the first Cambodian laboratory to obtain accreditation in accordance with the recognized international standard NF EN ISO 15189 by COFRAC in 2018.

Currently, a panel of over 150 tests and continuous development of new biomarkers are provided to all health professionals (including public and private hospitals, clinics, and nongovernmental organizations) working with LBM and all walk-in patients/clients.

The Laboratory is divided in to 4 different specialty areas :

- - Blood Biology Laboratory covers the area of Hematology, Coagulation testing, Biochemistry, Endocrinology and Immuno-serology, as well as Blood parasitology, Myelogram, FibroTest-ActiTest, Tumor markers dosage and semen analysis.

- - Medical Microbiology Laboratory is in charge of analysis in Bacteriology, Mycology and Parasitology.

- - Mycobacteriology Laboratory provides conventional and molecular diagnostic of Tuberculosis and Atypical Mycobacteria disease with adapted biosafety facilities (BSL2+ and BSL3).

- - Platform of Molecular Biology ensures molecular diagnostic of infectious diseases and rapid detection of resistance.

In order to provide a complete service to our clients, we collaborate with Laboratoire Cerba, which is the European leader in specialized clinical pathology, for very specialized biology tests.

Practical information

Those recommendations describes how to take bacteriological samples tested in Institut Pasteur du Cambodge. It is addressed to people taking bacteriological samples.

For others bacteriological samples or specific one, you may contact a biologist or get information at our reception.

For all the samples must be indicated systematically on the prescription:

- - First name, last name and date of birth

- - Time and date:
    - of the sampling
    - of the shipment to Institut Pasteur

1. - Identification of the medical doctor and of the nurse taking the sample

Analysis

Analysis of blood, urine and semen constituents

- Study of renal function (creatinine and creatinine clearance).
- Study of liver function (ALT, AST, GGT, PAL, Bilirubin, fibrotest).
- Study of carbohydrate metabolism (glucose, glycated hemoglobin), lipids (total cholesterol, LDL-cholesterol, HDL-cholesterol, triglycerides …), and anemia (serum iron, ferritin, transferrin, Vit B9, Vit B12, Haptoglobin, LDH, reticulocytes)

Serology (infectious and parasitology disease screening)

- Diagnosis of viral infections: HIV, hepatitis A, B and C, CMV, rubella
- Diagnosis of bacterial infections (syphilis, ASLO, typhoid fever)
- Diagnosis of parasitic infections (amoebiasis, toxoplasmosis, malaria).
- Follow-up of tumor markers (ACE, AFP, PSA, CA19-9, CA125 and CA15-3).
- Diagnosis of allergy (Eosinophil leukocytes, IgE).
- Study of endocrine dysfunction or therapeutic drug monitoring (thyroid hormones: TSHU, T3L, T4L; cortisol), sexual hormones (βhCG, FSH, LH, estrogen, progesterone, prolactin)

Hematology, Hemostasis, Immuno-hematology

- Count Blood Cells (red and white cells, haemoglobin, platelets).
- Diagnosis of leukemia.
- Haemostasis for the diagnosis of coagulation diseases or anticoagulant therapy monitoring (INR, aPTT, bleeding time).
- Diagnosis of haemoglobinopath.
- Blood grouping.
- Lymphocytes typing (CD4, CD8).
- Inflammatory markers (ESR, CRP, protein electrophoresis)

Microbiology: Bacteriology, Parasitology & Mycology

- Identification of bacteria, parasites or fungi, in miscellaneous samples (blood, pus, sputum, stool, urine, respiratory specimen…)
- Sexually transmitted infections diagnosis (gonococcal infection, Chlamydia, Mycoplasma and Herpes Simplex Virus).
- Antibiotic susceptibility testing by standardized methods.

Mycobacteriology

All these analysis are performed in biosafety level 2 and level 3 laboratories

- Diagnosis of tuberculosis and mycobacterium disease (direct smear, culture, GenXpert).
- Drug sensibility testing (first and second line anti-TB drug).
- Molecular biology (rapid detection of Mycobacterium Tuberculosis Complex and drug resistance and species identification of mycobacteria

Mycobacteriology

- Viral load of HIV, HCV and HBV.
- HIV and HBV resistance/ HCV genotyping.
- Diagnosis of encephalitis (H. influenzae type b, N. meningitidis, S. pneumoniae, S. suis)

Catalogue of Analysis

Quality Chart

Since its creation, the Medical Laboratory (LBM) of Institut Pasteur du Cambodge has been committed to a quality approach according to practical standards, especially the standard ISO 15189. In 2018, our Laboratory became the first Cambodian laboratory to obtain accreditation in accordance with the recognized international standard NF EN ISO 15189 by COFRAC.

Quality management at LBM aims at ensuring, for all customers:

- - Reliability and accuracy of test results
  - Reliability and accuracy of internal and external quality control
  - Continuous improvement to ensure consistent quality results
  - Data traceability
  - Confidentiality
  - Quality of the service
  - Satisfaction of clients
  - Reliable results interpretation based on the up-to-date recommendation
  - High level of technical skill of the staff
  - Technological development

Always at the forefront of a quality service, LBM has set up its Quality Management System in order to ensure the required quality standard at pre-analytical (sample collection and transportation), analytical (testing) and post-analytical (result delivery and interpretation) level, as well as the continual improvements to ensure consistent quality results. Our Laboratory develops continuously its human and technical resources by imposing strict quality standards. Our aim is to meet expectations and satisfactions of all clients and to collaborate with our partners with confidence based on the quality of the work that we perform daily.

Respectful of diversity and privacy, the laboratory management is also committed to ensuring that all biologists and employees respect the general principles of medical ethics and treat all of our patients fairly and without discrimination.

The organization of the laboratories is designed so that, in all working conditions, patient data is used only for strictly medical purposes. We do everything we can to keep the utmost confidentiality, whether at reception or at the reporting of results.

Knowing that analytical activities present a potential risk of contamination, we are committed to having a health and safety policy providing all the necessary safety both for patients and staff. After treatment, the samples are safely disposed of according to the legislation in force by approved channels. Appropriate procedures are in place to ensure that personnel handle human samples, tissue or residues safely.

The Laboratory checks the accuracy of the analysis results by performing quality control:

- - Internal: every analytical parameter is checked everyday
  - External: by taking part in external quality assessments (certificates of participation enclosed) in order to compare ourselves with other laboratories:
    – Association de Biologie Praticienne (ABP, France): hematology, serology, bacteriology, parasitology, mycology.
    – Bio-Rad (USA): biochemistry
    – Centre Toulousain pour le Contrôle de Qualité en Biologie Clinique (CTCB, France) : serology
    – Centre Lyonnais pour la PROmotion de la BIOlogie et du contrôle de QUALité (PRO.BIO.QUAL, France): hematology, coagulation, immunology
    – College of American Pathologists (CAP, USA): mycobacteriology
    – External Quality Assurance (EQUAS) : bacteriology
    – UK-NEQAS (Great Britain): CD3/CD4/CD8 lymphocytes
    – The Research Institute of Tuberculosis (Supranational Laboratory-Japan)/ National Center for Tuberculosis and Leprosy Control (NTP-Cambodia) (RIT/CENAT) : mycobacteriology
    – Quality Assessment and Standardization for Immunological Measures (QASI) : immunology
    – Roche External Quality Control Programm in Clinical Chemistry (REQC) : Biochemistry.

Our continuous quality improvement approach is supported by a quality management system combining several actions:

- - Writing and systematic diffusion to the staff of the quality documents (standard operating procedure (SOP), …)
  - interpretation of the results of the quality controls
  - Maintenance of the devices (internal maintenance and maintenance contracts with the suppliers approved by manufacturers)
  - Stock management of reagents (software used to avoid stock shortage and expiration of reagents)
  - Real time monitoring of the temperature of thermal enclosures (use of a specific software).

A plan for in-house training of the staff, as well as a technological development monitoring, allow us to maintain our high level of technical skill and to control our analytical tool.

We want to meet our patients’ expectations and to collaborate with our partners in a climate of confidence based on the quality of the work that we run daily.

Support to National Authorities

The following summarizes key support to Cambodian National Authorities during 2019 as part of our ongoing programs and projects.

- - With NCHADS, follow-up of HIV seropositives
  - Member of CENAT’s laboratory TWG for development of technical procedures guideline
  - Member of CENAT’s TWG on multi-drug resistant
  - Member of TWG on AMR with MOH
  - Provide technical support on quality with MOH

Teaching & Training

a. Continuing professional development training for staff

- - DU Qualité, accréditation et Audit, Université de Lille-II
  - Roche Quality Day 2019
  - Norme NF EN ISO 15189
  - “Breakthrough in Medical Technology” Congress
  - Massive open online course (MOOC), viruses and human cancers
  - MOOC, Quality Management of Medical Biology Laboratories
  - MOOC,The Role of Diagnostics in the Antimicrobial Resistance Response
  - MOOC, Epigenetics
  - Chemical and Biological Risk, Kalitech
  - Course in Antibiotic Resistance
  - Effective use of Kalidoc (document management tool)
  - Effective use of Maldi-TOF, Bruker
  - Cobas e411 refresher training
  - Introductory course in applied microbial genomics for public health and antimicrobial resistance

b. Internships

One of IPC’s main missions is to contribute to teaching and training activities. The medical laboratory has been proactive in the training of laboratory technicians and pharmacists from partner institutions in the fields of medical biology including haematology, immune-serology, biochemistry, microbiology and molecular biology. In 2019, the medical biology unit received 38 students for internship:

- - Pharmacists from UHS: 20
  - Laboratory Technicians from UHS & UP: 12
  - Kampong Cham Veterinary School: 6

c. Thesis supervision

One PhD candidate in health biology, University of Montpellier, studying the genetic determinants and evolution of drug resistance inMycobacterium tuberculosis in Cambodia, a high tuberculosis burden country

Supervising of students for their thesis project for the degree of doctor of pharmacy specializing in medical biology from UHS Cambodia.

- - Study of enterobacteriaceae producing extended-spectrum beta-lactamase at the Institut Pasteur in Cambodia (Étude desentérobactéries productrices de bêta-lactamase à spectre étendu à l’Institut Pasteur du Defended on August 2019.
  - Carbapenemase-producing bacteria, evolutionary monitoring at the Institut Pasteur in Cambodia (Bactéries productrices de carbapénémases, suivi évolutif à l’Institut Pasteur du Cambodge). Thesis is
  - Study of mutations in genes linked to the resistance of Mycobacterium tuberculosis isolated from patients in Cambodia (Étude des mutations des gènes liés à la résistance de Mycobacterium tuberculosis isolées des patients au Cambodge). Thesis is ongoing.

Jury member to evaluate a thesis for the degree of doctor of pharmacy specializing in medical biology from UHS.

- - Surveillance of the presence of leptospire and Hantavirus carried by rodents in four provinces of Cambodia. Defended on March (Surveillance de la présence de leptospire et d’hantavirus portés par les rongeurs dans quatre provinces du Cambodge). Defended on March 2019.

d. Participation in the setting up of master’s course at UHS

- - Co-coordinator of bacteriology module for Master’s years one and two in life sciences and international track in infectious disease studies in Cambodia. This degree offered by UHS Cambodia, the University of Paris- Saclay and IPC, Cambodia is an advanced level program and is intended as a continuation of the French bachelor’s degree in life
  - Co-coordinator of bacteriology module for master’s in medical biology,

Projects – 2020

Antibiotic resistance in the human, animal and environment interface in an intervention in Cambodia: ARCAHE

In recent years, antibiotic resistance rate has become critical since multiple and extended resistant bacteria have become more prevalent. Even if we know that the current antibiotic resistance crisis is due to the massive use, overuse and misuse of these drugs, in many countries including Cambodia that have significant weakness in antibiotic usage regulation, still very little data are available and nosurveillance system exists. With this background, we submitted a project that aims to identify the sources of emergence and spread ofresistant bacteria in Cambodia using a “One Health” approach. This project will have a huge impact since for the first time, it will explore thespread of antibiotic resistance in humans, animals and environment in Cambodia. The expected results will represent the baseline for thesetting up of a surveillance system, will allow stakeholders to implement efficient control strategies. The initiative will be done with IRD, Calmette, and Battambang Hospitals.

Team leader: Sokleaph CHENG

Funding: FSPI, French Embassy in Cambodia

AVIESAN- South initiative on antimicrobial resistance

The objective of this activity is to setting up a research network in countries in the south (Sub-Saharan Africa and South-East Asia) to develop research projects related to antimicrobial resistance. Four countries were identified (Cambodia, Madagascar, Ivory Coast and Burkina Faso) and the visit of the Aviesan south delegation to those countries were started since late 2019. The priorities research themes on AMR were will be identified by each countries and will be presented to Aviesan SUD. One or two common research themes willbe selected, pilot studies will be conducted to test its feasibility of each research project and the full projects will be applied for funding.

Team leader: Sokleaph CHENG

Funding: Aviesan SUD (Aviesan south)

Pertussis genotyping from nasopharyngeal samples collected in Cambodia (Perilic WP1 – ancillary study)

The objective of this ancillary study being done in partnership with IP Paris is to characterize the main relevant genes carried by B.pertussis strains collected in Cambodia as part of the PERILIC study, directly using the primary nasopharyngeal samples. In particular, macrolides resistance and pertactin expression will be assessed and virulence gene alleles (including prn, ptxa, ptxc, ptxp, fim2, fim3) will be determined by genotyping.

Team leader: Sokleaph CHENG

Funding: Institut Pasteur- Paris

New AntiMicrobial Peptides (AMP)

The general objective of the study is to evaluate the antibacterial activity of a new set of eight synthetic AMP against clinical isolates ofrelevant bacteria identified by GLASS. The antibacterial activity of AMP against relevant bacteria involved in human infections in Cambodia (Acinetobacter spp., Escherichia coli, Klebsiella pneumoniae, Salmonella spp., Shigella spp., Staphylococcus aureus) will beevaluated. Experiments will start in May 2020 by a master’s of medical biology student).

The specific objectives are to set up antibacterial activity testing using broth microdilution method for the eight AMP on E. coli reference strain (ATCC 25922), and to measure the susceptibility of relevant resistant bacteria identified by GLASS against the eight new AMP by comparing with sensitive bacterial strain. Partners include LMI-DRISA and MIVEGEC.

Team leader: Mallorie HIDE (IRD – LMI DRISA)

Funding: LMI-DRISA, IPC

Potential colistin resistance in multidrug-resistant Enterobacteriaceae

The current antibiotic of last resort for the treatment of infections in humans caused by multidrug-resistant enterobacteria is often colistin. It belongs to the antimicrobial class designated polymyxins which originates from the organism Paenibacillus polymyxa. In Cambodia,colistin is one of the most commonly used antibiotics on small- scale urban pig farms and is used on poultry farms in a packaged amoxicillin-colistin mixture for respiratory and gastrointestinal infections. On pig farms, 20% of phenotypic resistance to colistin have beenidentified. A few colistin resistance genes (mcr like, pmrA, pmrB, phoP, phoQ, mgrB, and pmrD genes) have been detected from both multidrug‐resistant Escherichia coli (EC) and Klebsiella pneumonia (KP) isolated from humans, poultry and pigs. Despite this, colistin resistance in multidrug-resistant Enterobacteriaceae isolated from patient is poorly known in Cambodia. With this background, our main objective is to explore colistin resistance among both carbapenemase and extended‐spectrum Beta-Lactamase (ESBL) isolated fromhumans.

Experiments (in vitro testing, sequencing) will start in May 2020, working with a master’s in life sciences and international health student specializing in infectiology-biology of infectious diseases. Implementation will be done in collaboration with LMI DRISA and MIVEGEC.

Team leader: Mallorie HIDE (IRD – LMI DRISA)

Funding: LMI-DRISA, IPC

Genetic diversity and resistance status of Burkholderia pseudomallei strains responsible for melioidosis in Cambodia

Burkholderia pseudomallei is an environmental gram-negative bacteria, transmitted to humans by contaminated soil and surface water (percutaneous inoculation, inhalation, or ingestion) and most of cases occur during rainy and humid months. Melioidosis, due to B.pseudomallei, is endemic in Cambodia and the majority of cases are in children (>60%) with head and neck infections whereas adults mostly present pneumonia and/or sepsis with increase factors such as diabetes mellitus, renal diseases, chronic lung disease,thalassemia, alcohol consumption, male sex and occupational exposure. Concerning antibiotic resistance, most Burkholderia contain a modified lipopolysaccharide that causes intrinsic polymyxin resistance and B. pseudomallei exhibits resistance to diverse antibiotics including first and second generation cephalosporins, penicillins, macrolides, and aminoglycosides. Ceftazidime and clavulanic acid-resistant strains have been described and others continue to be identified. No genetic data are available on Burkholderia in Cambodia and in this context, our objective is to explore the genetic diversity and resistance status of Burkholderia strains responsible for melioidosis, isolated and cryoconserved at the Medical Laboratory since 2011 (n=100). All the bacterial samples will be cultured foridentification using Bruker Maldi Biotyper and sequence to explore both genetic diversity and resistance genes.

Experiments (Maldi-tof identification, AST, genetics) will start in May 2020. A Master’s of live sciences and international health studentspecializing in the biology of infectious diseases will work on this study under supervision of IPC. Partners in this work will be LMI-DRISA, and MIVEGEC

Team leader: Mallorie HIDE (IRD – LMI DRISA)

Funding: LMI DRISA, IPC

Establishing an AMR bacteriology research focus in IPC

During his postdoctoral position, K. Vandelannote will define genomic characteristics and population structure of pathogen specificoutbreaks using whole genome sequencing and model transmission networks using clinical and genomic epidemiological data.

Projects – 2019

PTR on aspergillosis and aspergillus in Cambodia

This PTR received ethical approval from the Cambodian National Ethics Committee for Health Research on December 26, 2017. It is worth noting that due to ethical, financial, clinical and practical constraints, some adaptations were necessary and a focus on Aspergilloma was decided (new acronym: “AspA” for aspergilloma in Cambodia). A sponsorship transfer from IP Paris to Institut Pasteur Cambodia was accepted. The main objective of the project was the setup of the first mycology lab in the Pasteur Institute International Network. This objective was fully accomplished. Furthermore a clinical research assistant (Mala Sim) and a technician (Sreyroth Lim) were recruited to help the patient inclusion process, the monitoring of the study and lab experiments. The patient recruitment was finalized in early October 2018 with 645 included patients. Of these, 53.2% were men (mean age of 57.65 years +/- 16.18) and 46.8% were women (mean age 59.15 years +/- 15.59). Patelia ELISA® (gold standard) was positive for 14.54% of the patients tested (81/557). Eight recombinant proteins (Gel1, Crf1, Cat1, and 18Kd, Sod1, A97, H41 and H70) were tested for diagnosis purposes and only two allowed the differentiation between positive and negative serum control. The recombinant protein A97 showed promising results with arelative sensitivity of 72% and a specificity of 93%.The direct examination of the sputum was positive for 37.7% (210/557) of the patient.Four hundred and eight strains were isolated from patient and 63 from the hospitals environment.

Most of the strains belonged to the A. niger / A. tubingensis clade then A. fumigatus and A. flavus. Interestingly, the mutation TR34/L98H/T364A was recorded in Cyp51A gene of one A. fumigatus strain. Antifungal assays showed this mutation is responsible for a 16-fold increase of the minimal inhibitory concentration for itraconazole and thus indicate a high resistance phenotype (MIC > 2 µg/ml instead of 0.12-1 µg/ml for Aspergillus fumigatus wild type KU80). Descriptive statistics of the included patient reveal a high proportion of productive cough longer than one month (78%), weight loss (74.4%), dyspnea longer than one month (68%) and hemoptysis (47.9%). Acomparison between the whole sample, TB negative patients and TB negative and Platelia positive patients highlight no statistical difference for diagnosis based on direct examination in the sputum and fungal culture but a tendency to have more positive fungal culture when TB negative and Platelia positive. Concerning these three populations, no statistical difference was found for Platelia and Ct-scan results. In the same way, no statistical difference was found between Platelia negative and positive patients nor Ct-scan positive and Ct-scan negative patients. The main limitations of this study were the poor quality of the sputum (58% of the sputum below the Murray-Washington class 3, the high number of patients with unknown tuberculosis status (24.4%) and the exclusion of patients due to absent Ct-scan (Platelia positive). We hope these first results on aspergilloma diagnosis will help physicians to improve health care of patient and particularly concerning the management of tuberculosis sequelea. An « Aspa day » was organized near the close of the project for scientific restitution and acknowledgment and recognition of all collaborators. The PTR was closed at the end of June 2019 following a last no-cost extension. The project was jointly implemented by three Phnom Penh Hospitals- Calmette, Khmer-Soviet Friendship, and Kossamak. An oral presentation was done in Paris by Dr. Chhorn Sotheary in January 2020 during the 24th Pneumology Congress. Thepublication is in progress and will be submitted in an international scientific journal as soon as possible.

Team leader: Y. Caron

Funding: IP – PTR

Outbreak of trichinellosis in September 2017 in Kampong Thom Province

On September 26, 2017, an important outbreak (33 cases including 8 deaths) of trichinellosis (Trichinella spp.) in Central Kampong ThomProvince Hospital was reported. Trichinellosis was then diagnosed in Phnom Penh in two hospitals: Calmette and Preah Ket Mealea Hospitals. A proposal to investigate the parasite with the help of the Trichinella Reference Laboratory (Cochin Hospital, Paris, France) tothe communicable disease control department of the MOH was accepted. We accessed to the complete medical records (clinical symptoms, diagnosis, medical history, treatment, outcome etc.) of each hospitalized patient in the third hospital experiencing this outbreak. Very few scientific data exists in Cambodia on this zoonotic disease and the molecular characterization of the strain involved could be interesting for the management of this important public health issue. A publication entitled “First description of Trichinellapapuae involved in a human outbreak in central Kampong Thom Province in Cambodia » was submitted to the journal “Emerging Infectious Disease”; the review process is ongoing.

Team leader: Y. Caron

Funding: IP Cambodia

International joint laboratory «Drug Resistance in Southeast Asia» (LMI DRISA)

In collaboration with the IRD, IPC has hosted an engineer specialized in molecular biology since August 2018. Dr. Mallorie HIDE will be based in LBM for at least three years within the LMI DRISA framework. She works on molecular epidemiology and drug resistance inMycobacterium tuberculosis and other bacteria in Cambodia as well as on the application of antimicrobial peptides as alternatives toantibiotics.

Laboratory on bacteriology research (LRB)

Within the LBM, an analytic platform dedicated to research in bacteriology has been set up by M. Hide thanks to the financial supportprovided by IPC and LMI DRISA. For both routine activity and research purpose, IPC bought a Bruker biotyper Maldi-tof, currently availablein the LRB. For the molecular characterization of tuberculosis isolates, the automated spoligotyping has been set up using Luminexhardware available on the PRR -ASIA platform.

Genetic determinants and evolution of drug resistance in Mycobacterium tuberculosis in Cambodia, a high tuberculosis burden country

The overall objective of this project is to understand the emergence, spread and evolution of antibiotic resistance in Mycobacterium tuberculosis in Cambodia by using genetic characterization of MTB isolates. This is a three year study started in September 2016. By 2018, a total of 473 MTB isolates with known drug susceptibility testing (DST) for fragmented and loop primer ligated dsRNA sequencing (FLDs)(Isoniazid, rifampicin, streptomycin and ethambutol), collected between 2012 and 2015 were included in the study. Among them, 143 were sensitive to FLDs and 326 were resistant to at least one FLD. These isolates were genetically characterized using sequencing of genes involving in the resistance of MTB to first and second line anti-TB drugs and genotyping method. In 2019, the genetic data wereobtained for 404 isolates and included in the analysis. Of these 278 were resistant to at least one FLD and 126 were FLD sensitive. Of drug resistant isolates, 113 (28%) were mono-resistant (resistant to only one FLDs), 44 (10.9%) were poly-resistant (resistant to more than one FLDs but not to isoniazide and rifampicine at the same time) and 115 (41.2%) were MDR. The resistance to INH and STM were the most commonly found among our isolates, representing 52.0% (210/404) and 42.3% (171/404), respectively. Rifampicin resistance was detected in 140 (34.7%) isolates, of them 17 (4.2%) were mono-rifampicin resistant. Ethambutol resistance was less common, observed mostly among MDR isolates (48/121). The isolates resistant to all four FLDs, defined as “quadruple resistant”, were found in 9.9% of all isolates. The demographic and clinical data linked to the 404 MTB clinical isolates were collected from the patient recorded.The median age was 44.7 and ranged from 15 to 86 years old, with 63% being male. The majority of patients (97.5%, 318/326) had pulmonary TB. Among 163 patients with known TB treatment history, most study subjects were re-treatment TB cases 134 (82.2%) and 7 (4.3%) patients were people living with HIV. At the time samples were collected for culture, 65.8% (206/313) were smear positive.

43-spacers spoligotyping results were available for 403 isolates and revealed eight families and 22 sub-families. Beijing and EAI familieswere predominant, accounting for 46.4% and 43.4%, respectively. The 24 loci MIRU-VNTR typing of 404 M.tb isolates allowed todistinguished 360 patterns, including 307 unique patterns, 34 patterns shared by 78 isolates (ranging from 2 to 4 isolates per pattern) and 19 mixed genotype patterns (double alleles in at least one locus). A dendrogram was constructed based on both spoligotyping and MIRU results of 384 M.tb isolates and 31 clusters were identified which suggest a recent transmission among patients. In deep analysis are being conducted and two publications are in preparation (one focus of highly resistant isolates and other focus on genetic diversity and their link to drug resistance among all isolates). The data from this project is being used in one PhD thesis and one resident in medical biology for a thesis, and will be used in a second DES thesis. The project is a collaboration with CENAT the Ministry of Education, Youth, and Sport, and the PHC Lotus Project of NIHE.

Team leader: Sokleaph CHENG

Funding: LMI-DRISA, IRD, French Embassy in Cambodia

In 2019, IPC submitted five projects to different calls for application of which was accepted for funding by the Fonds de solidarité pour les projets innovants (FSPI), Embassy of France in Cambodia.