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Supranational TB Reference Laboratory in Buenos Aires, Argentina
Address:
David Avendaño (davendaño@anlis.gov.ar)
Norberto Fabian Simboli (nsimboli@hotmail.com; nsimboli@anlis.gov.ar)
Mycobacterias Divition
Av. Vélez Sarsfield 563 CABA (C1282AFF)
Buenos Aires City Argentina
Description
The responsibilities of our division at a National level are as a Reference Laboratory for the TB laboratories network and as a Reference Laboratory for mycobacteriosis.
We are mainly focused on the study of tuberculosis, and it provides technical assistance to National Control Programs of Tuberculosis and Human Retrovirus.
Our laboratory’s main activities are:
Reference diagnosis
Production and distribution of reagents
Surveillance of resistance anti-tuberculosis drugs
Monitoring of Mycobacteriosis
Establishing of rules and standards
Training of human resources
Validation of methods
Quality management and biosafety
Structure and organization
The mycobacteria Laboratory belongs to the National Institute of Infectious Diseases INEI- ANLIS “Carlos G Malbran”. It integrates, sharing with E Coni National Lab, the National TB Laboratory Network and it supports the National TB Program. It includes about 850 labs classified in different levels of complexity, 705 primary health units, 125 at an intermediate level and 25 at a provincial level. Each level has different tasks.
The lab covers an area with three different sections, the upper floor includes the BSL3 lab, the computers office and the washing and storage area, the lower floor includes the clean area with offices and 3 labs. One of the labs is separated from the rest as it is a dirty section used for molecular biology.
Services provided by the SRL
Our laboratory centralizes the execution of some techniques as detection of adenosine deaminase in pleural liquid, identification of the mycobacteria no tuberculous at specie level and determination of the profile of sensitivity to first, second line and alternatives drugs. We have been keeping records of cases with multi-drug resistance and extended-resistance tuberculosis since 2003, we define the sensitivity profile of second-line drugs of these strains, and monitor the transmission of strains causing outbreaks and we also participate in the national commission of experts in re-treatment. We keep up to date with the frequency of mycobacterial non tuberculosis infection, and define its sensitivity profile to potential drugs and characterize the mycobacteria outbreaks.
Diagnosis of pleural TB:
Adenosine deaminase assay in pleural liquid
Identification of mycobacterias at level species:
Lateral immune-chromatography technique
PCR “in house” to detect IS6110
PRA hsp65-PCR restriction enzyme analysis of the hsp65 gene
ALPACTB: Pattern analysis from PCR “in house” based in different chromosomal locus to differentiate species of MTB Complex
MALDI TOF: Protein patterns using the matrix-assisted laser desorption/ionisation time-of-flight
Phenotypical methods
Sequencing of genes 16S, rpoB, sodA
Susceptibility profile:
MAS-PCR: Multiplex allele specific PCR “in house” to detect rifampicin and isoniazid
Proportional method in Lowenstein Jensen to first and second line drug
Proportional method in MGIT 960 with TB eXiT software to first, second and alternative drugs
Wayne method
Minimal inhibitory concentration in liquid medium to alternative drugs in XDR TB
Sequencing of gene rpoB
Minimal inhibitory concentration in liquid medium to potential drugs in micobacteriosis
Genotyping method
Mycobacterium tuberculosis
Spoligotyping
DRE-PCR: Double Repetitive Element PCR
MIRU-VNTR typing based on 15 and 24 locis
Rapidly Growing Mycobacterium Species
ERIC method: Enterobacterial repetitive intergenic consensus PCR
Current links and partnerships outside of SRLN
The two last years, we have had links and partnerships out our country with :
Laboratory of Microbiology, Department of Biochemistry & Microbiology, Ghent University, Ghent, Belgium.
Division of Infectious Disease Control, Norwegian Institute of Public Health, Oslo, Norway
Department of Preventive Medicine, School of Medicine, Autonoma University of Madrid, Madrid, Spain
Molecular Genetics & Biotechnology, Corporación CorpoGen, Bogotá, Colombia
Laboratory of Molecular Biology Applied to Mycobacteria, Oswaldo Cruz Institute, Rio de Janeiro, Brazil
Biological Science National School, Instituto Politécnico Nacional, Distrito Federal, México
Department of Medicine, Universidad Federal de Sergipe (UFS), Aracaju, SE, Brazil
Additional information:
Research activities
Grant-aided
- Identification of bioequivalent concentrations of levofloxacin and moxifloxacin between Löwenstein-Jensen and liquid medium. PAHO/WHO grant, 2015-2016.
INEI- ANLIS regular budget
- Evaluation of specific allele multiplex PCR (MAS-PCR) for MDR-TB detection.
- Evaluation of MALDI-TOF MS methodology for identification of clinically-relevant non tuberculouos mycobacteria.
- Integration of quality-controlled MIRU-VNTR methodology to our Mycobacterium tuberculosis genotyping armamentarium.
- Development of an improved multiplex PCR (MAS-PCR) for detection of M. tuberculosis fluoroquinolone resistance.
- Sequencing of the rpoB gene for resolving discrepant results of phenotypic and genotypic methods for rifampicin resistance detection.
- Validation of the resazurin microplate method for MIC assessment of drugs potentially useful for treatment of mycobacteriosis.
National collaborations
- Characterization of XDR Mycobacterium tuberculosis strains from Argentina by whole-genome sequencing. Post-doc project in collaboration with Plataforma Bioinformática Argentina, Instituto de Cálculo - Facultad de Ciencias Exactas y Naturales. UBA. 2016- continues
- Interaction of the macrophage with clinical isolates of Mycobacterium tuberculosis and relationship with their epidemiological success. Partner: IMEX-CONICET, Academia Nacional de Medicina. MinCyT project, 2012-continues.
- M. tuberculosis manipulation of macrophage metabolism as an immune evasion strategy. Mincyt Agency, Argentina PICT 2015-0055. 2016- continues.
International collaborations
- Impact of the HIV infection on the evolution of two conspicuous MDR M. tuberculosis outbreak strains from Argentina. Partners: Norwegian Institute of Public Health, University College London and Imperial College, UK. 2014- continues.
- Whole-genome sequencing for characterization of clinical isolates of M. bovis BCG obtained from disseminated BCG infection. Partner: Oswaldo Cruz Institute, Brazil. 2015- continues.
- A novel transnational strategy to control high-risk tuberculosis transmission events. Partner: Hospital Universitarion Gregorio Marañon, Madrid, Spain, and others. ERANET-LAC EC call 2016 (grant project under evaluation).
- Elucidating the mechanisms of antimicrobial evasion by Mycobacterium tuberculosis. Partner: Ghent University, Belgium. Bilateral cooperation Mincyt Argentina-FWO Belgium FW/13/03 2014-2016.
- Micobacterias: Carácter patógeno y relación con el entorno. Partners: Universidad Autónoma de Madrid and others. Proyect CEAL AL/2015-22. 2015-2016 (continues).
Publications
(last 5 years)
- Ritacco V, Iglesias MJ, Ferrazoli L, Monteserin J, Dalla Costa ER Cebollada A, Morcillo N, Robledo J, Araya P, de Waard J, Aristimuño L, Díaz R, Gavin P, Imperiale B, Rossetti ML, Jiménez MS, Martin C, Barrera L, Samper S. Conspicuous multidrug-resistant Mycobacterium tuberculosis cluster strains do not trespass country borders in Latin America and Spain. Infect Genet Evol 2012;12: 711–7. IF 3.3
- Basile JI, Geffner LJ, Romero MM, Balboa L, Sabio Y García C, Ritacco V, García A, Cuffré M, Abbate E, López B, Barrera L, Ambroggi M, Alemán M, Sasiain MC, de la Barrera SS. Outbreaks of Mycobacterium tuberculosis MDR Strains Induce High IL-17 T-Cell Response in Patients With MDR Tuberculosis That Is Closely Associated With High Antigen Load. J Infect Dis. 2011;204:1054-64. IF 6.0
- Ritacco V, López B, Ambroggi M, Palmero D, Salvadores B, Gravina E, Mazzeo E, National TB Laboratory Network, Imaz S, Barrera L. HIV infection and geographically bound transmission of drug-resistant tuberculosis, Argentina. Emerg Infect Dis. 2012;18: 1802-10. IF 7.3
- Abbate E, Vescovo M, Natiello M, Cufré M, García A, Gonzalez Montaner P, Ambroggi M, Ritacco V, van Soolingen D. Successful alternative treatment of extensively drug-resistant tuberculosis in Argentina by a combination of linezolid, moxifloxacin and thioridazine. J Antimicrob Chemother. 2012;67:473-7. IF 5.3
- Yokobori N, López B, Geffner L, García CS, Schierloh P, Barrera L, de la Barrera S, Sakai S, Kawamura I, Mitsuyama M, Ritacco V, Sasiain MC. Two genetically-related multidrug-resistant Mycobacterium tuberculosis strains induce divergent outcomes of infection in two human macrophage models. Infect Genet Evol 2013;16:151-6. IF 3.3
- Eldholm V, Monteserin J, Rieux A, Lopez B, Sobkowiak B, Ritacco V, Balloux F. Four decades of transmission of a multidrug-resistant Mycobacterium tuberculosis outbreak strain. Nat Commun. 2015 May 11;6:7119. IF 10.7
- Romero MM, Basile JI, Correa Feo L, Lopez B, Ritacco V, Alemán M. Reactive oxygen species production by human dendritic cells involves TLR2 and dectin-1 and is essential for efficient immune response against mycobacteria. Cell Microbiol. 2015 Dec 28. [Epub ahead of print]. IF 4.9
- Monteserin J, Paul R, Lopez B, Cnockaert M, Tortoli E, Menéndez C, García MJ, Palomino JC, Vandamme P, Ritacco V, Martin A. Multiple approach for identification of clinically infrequent non tuberculous mycobacteria in Argentina. Int J Tub Lung Dis (in press 2016). IF 2.3
Trainings
We are making the training in internships in different topics according to demand of the laboratories
Annually we are organizing one or two courses in different activities according to demand the tuberculosis laboratories network
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