Publications by authors named "Maria Alice da Silva Telles"

12 Publications

  • Page 1 of 1

Genetic diversity of the Mycobacterium tuberculosis Beijing family in Brazil and Mozambique and relation with infectivity and induction of necrosis in THP-1 cells.

Tuberculosis (Edinb) 2015 Jun 25;95 Suppl 1:S190-6. Epub 2015 Feb 25.

Laboratory of Molecular Biology Applied to Mycobacteria, Instituto Oswaldo Cruz, Rio de Janeiro, Brazil. Electronic address:

Introduction: The success of the Mycobacterium tuberculosis Beijing (MtbB) lineage in different geographical regions has been attributed to high transmission, increased virulence, drug resistance and rapid adaptation to the host. In some countries of secondary MtbB dispersion like South Africa and Peru, rising prevalence of the Beijing strains is registered. However, in neighboring countries to affected regions such as Mozambique and Brazil, respectively, the prevalence of these strains is still low and this could be due to biological particularities of the circulating MtbB strains and/or differentiated host susceptibility.

Objective: To characterize genetically and phenotypically MtbB strains isolated in Brazil (n = 8) and Mozambique (n = 17).

Methods: This is a descriptive study of genotypes of the MtbB isolates, determined by spoligotyping, MIRU-VNTR typing, analysis of the IS6110 copy number in the NTF region and screening for mutations in mutT2, mutT4, rpoB, katG and pks 15/1 genes. Virulence-associated properties of the studied isolates were verified in the in vitro model of infection of human THP-1 cells.

Results: The genotypes defined by the 24VNTRs were distinct for all isolates included in this study and presented an HGDI of 0.997. The VNTR patterns with seven copies of MIRU26 and seven copies of QUB26, representative for the previously described MtbB genotype B0, dominant in Russia, were detected in 38.5% of the studied isolates. In addition, all isolates presented RD105 deletion and a 7 bp insertion in pks15/1 gene. Almost all tested strains belonged to the RD181 sublineage, with the exception of two strains from Mozambique of RD150 sublineage. Combined analysis of the NTF region integrity and mutations in mutT genes showed that 62.5% and 47% of isolates obtained in Brazil and Mozambique, respectively, were of the ancestral genotype. The virulence index of the ancient isolates, evaluated in the THP-1 cells, was significantly lower than that of the modern genotype group.

Conclusions: These data demonstrate genotype particularities of the Beijing strains isolated in Brazil and Mozambique, two countries of low prevalence of the MtbB lineage in local Mtb populations. In contrast to the neighboring countries with high prevalence of the MtbB strains of modern sublineage, significant proportions of the isolates obtained in Brazil and Mozambique were presented by the strains of the ancient sublineage. Our data suggest that lower virulence of the ancient strains, compared with the modern strains, could be involved in the slow spread of the MtbB strains in some regions.
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http://dx.doi.org/10.1016/j.tube.2015.02.025DOI Listing
June 2015

Bottlenecks and recommendations for the incorporation of new technologies in the tuberculosis laboratory network in Brazil.

J Bras Pneumol 2012 Nov-Dec;38(6):766-70

Instituto Adolfo Lutz, São Paulo, SP, Brasil.

The World Health Organization (WHO) has recently recommended new technologies for the diagnosis of tuberculosis. The WHO recommendations include the development of a strategic plan for bringing the network up to grade; investment in supervision and quality control; and implementation of a system of laboratory environmental management. Without those measures having been taken, no new technology can be effectively incorporated. We surveyed the tuberculosis laboratory network in Brazil in order to identify possible bottlenecks for the incorporation of new technologies. We identified a lack of resources allocated to supervision and quality control; a low number of requests for cultures; a lack of effective laboratory information systems; and a lack of awareness regarding the future infrastructure needs of the laboratory network at the municipal level.
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http://dx.doi.org/10.1590/s1806-37132012000600013DOI Listing
October 2013

Susceptibility of Mycobacterium tuberculosis to first-line antimycobacterial agents in a Brazilian hospital: assessing the utility of the tetrazolium (MTT) microplate assay.

Mem Inst Oswaldo Cruz 2010 Aug;105(5):661-4

Instituto Adolfo Lutz, Secretaria de Saúde do Estado de São Paulo, São Paulo, Brasil.

We conducted a cross-sectional, hospital-based study between January 2006-March 2008 to estimate the resistance of Mycobacterium tuberculosis to first-line drugs in patients with tuberculosis at a Brazilian hospital. We evaluated the performance of the [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide] (MTT) microplate assay compared with the Bactec-MGIT 960 system for mycobacteria testing. The prevalence of resistance in M. tuberculosis was 6.7%. Multidrug-resistance [resistance to rifampicin (RMP) and isoniazid (INH)], INH-resistance and streptomycin (SM)-resistance accounted for 1%, 3.8% and 3.8% of all resistance, respectively, and all isolates were susceptible to ethambutol (EM). The resistance was primary in four cases and acquired in three cases and previous treatment was associated with resistance (p = 0.0129). Among the 119 M. tuberculosis isolates, complete concordance of the results for INH and EM was observed between the MTT microplate and Bactec-MGIT 960TM methods. The observed agreement for RMP was 99% (sensitivity: 90%) and 95.8% for SM (sensitivity 90.9%), lower than those for other drugs. The MTT colourimetric method is an accurate, simple and low-cost alternative in settings with limited resources.
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http://dx.doi.org/10.1590/s0074-02762010000500010DOI Listing
August 2010

Cord factor detection and macroscopic evaluation of mycobacterial colonies: an efficient combined screening test for the presumptive identification of Mycobacterium tuberculosis complex on solid media.

J Bras Pneumol 2009 Dec;35(12):1212-6

Instituto Adolfo Lutz, São Paulo, Brazil.

Objective: The rapid differentiation between Mycobacterium tuberculosis and nontuberculous mycobacteria is fundamental for patients co-infected with tuberculosis and HIV. To that end, we use two methods in our laboratory: detection of cord factor and PCR-restriction enzyme analysis (PRA). The objective of this study was to evaluate the accuracy of a screening test on solid medium as a rapid method for the presumptive identification of M. tuberculosis complex, considering costs and turnover time.

Methods: A total of 152 strains were submitted to a combined screening test, consisting of the detection of cord factor under microscopy (Ziehl-Neelsen staining) and evaluation of the macroscopic aspect of colonies, as well as to PRA, which was used as the gold standard. Costs were estimated by calculating the price of all of the materials needed for each test.

Results: The overall accuracy of cord factor detection alone was 95.4% (95% CI: 90.7-98.1%), and that of the combined screening test was 99.3% (95% CI: 96.4-100%). Cord factor detection costs US$ 0.25, whereas the PRA costs US$ 7.00. Results from cord factor detection are ready in 2 days, whereas PRA requires 4 days to yield results.

Conclusions: The presumptive identification of M. tuberculosis using the macroscopic evaluation of colonies combined with cord factor detection under microscopy is a simple, rapid and inexpensive test. We recommend the combined screening test to rapidly identify M. tuberculosis in resource-poor settings and in less well-equipped laboratories while awaiting a definite identification by molecular or biochemical methods.
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http://dx.doi.org/10.1590/s1806-37132009001200008DOI Listing
December 2009

III Brazilian Thoracic Association Guidelines on tuberculosis.

J Bras Pneumol 2009 Oct;35(10):1018-48

New scientific articles about tuberculosis (TB) are published daily worldwide. However, it is difficult for health care workers, overloaded with work, to stay abreast of the latest research findings and to discern which information can and should be used in their daily practice on assisting TB patients. The purpose of the III Brazilian Thoracic Association (BTA) Guidelines on TB is to critically review the most recent national and international scientific information on TB, presenting an updated text with the most current and useful tools against TB to health care workers in our country. The III BTA Guidelines on TB have been developed by the BTA Committee on TB and the TB Work Group, based on the text of the II BTA Guidelines on TB (2004). We reviewed the following databases: LILACS (SciELO) and PubMed (Medline). The level of evidence of the cited articles was determined, and 24 recommendations on TB have been evaluated, discussed by all of the members of the BTA Committee on TB and of the TB Work Group, and highlighted. The first version of the present Guidelines was posted on the BTA website and was available for public consultation for three weeks. Comments and critiques were evaluated. The level of scientific evidence of each reference was evaluated before its acceptance for use in the final text.
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http://dx.doi.org/10.1590/s1806-37132009001000011DOI Listing
October 2009

Rapid detection of resistant tuberculosis by nitrate reductase assay performed in three settings in Brazil.

J Antimicrob Chemother 2009 Oct 11;64(4):794-6. Epub 2009 Aug 11.

Regional Laboratory Sorocaba, Brazil.

Objectives: To evaluate nitrate reductase assay (NRA) efficacy for streptomycin, isoniazid, rifampicin and ethambutol susceptibility testing of Mycobacterium tuberculosis strains.

Methods: Results were generated by three laboratories: the Instituto Adolfo Lutz (IAL) Mycobacteria Reference Laboratory and two IAL Regional Laboratories in Santo André and Sorocaba, São Paulo State, Brazil. One hundred and twenty M. tuberculosis strains were simultaneously tested using NRA and the proportion method (PM), while 117 strains were tested using both NRA and BACTEC MGIT 960 (M960).

Results: Repeatability analysis of NRA results showed rates of 100% for isoniazid and ethambutol and 97% for streptomycin and rifampicin susceptibility detection, representing substantial agreement. McNemar testing of the data also indicates that NRA and PM, as well as NRA and M960, do not differ significantly. On average, NRA results were available after 10 days.

Conclusions: The data demonstrate that NRA is reliable for susceptibility testing of isoniazid and rifampicin, the two most important drugs for the treatment of tuberculosis. In addition, the reduction in the time necessary to obtain susceptibility results is of fundamental importance.
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http://dx.doi.org/10.1093/jac/dkp284DOI Listing
October 2009

Maintenance of Mycobacterium tuberculosis on glass beads.

Ann Clin Lab Sci 2009 ;39(1):51-4

Instituto Adolfo Lutz, São Paulo, Brazil.

The intent of this study was to estimate the shelf life of Mycobacterium tuberculosis strains, and to observe the loss of viability in some of them from year to year. From 2000 to 2004, 10,015 cultures of M. tuberculosis were preserved by freezing on glass beads at -70 degrees C. With the expectation that the loss of viability might be around 5-10%/yr of storage, 730 strains were analyzed in order to establish the prevalence of recovery within a 5% margin of error. This study shows that 94% of the strains preserved at -70 degrees C on glass beads could be recovered within 30 days. The recovery rates for drug-susceptible and drug-resistant strains showed no significant differences. The growth rates and the number of strains that showed abundant growth before the 30th day of incubation represent important features, since the subculture of a strain preserved for future use ought to quickly produce abundant growth in order to avoid misinterpretation of the tests. Our experience indicates that storage of M. tuberculosis on glass beads at -70 degrees C is a suitable procedure for an active culture collection in a public health laboratory like ours, where maintenance of M. tuberculosis cultures is a complementary activity and must be quick, practical, effective, and economical.
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March 2009

Identification of non-tuberculous mycobacteria from the Central Public Health Laboratory from Mato Grosso do Sul and analysis of clinical relevance.

Braz J Microbiol 2008 Apr 1;39(2):268-72. Epub 2008 Jun 1.

Laboratório Central de Saúde Pública de Mato Grosso do Sul , Mato Grosso do Sul, MS , Brasil.

Non-tuberculous mycobacteria isolated at the Central Public Health Laboratory from Mato Grosso do Sul in 2003 and 2004 were identified by conventional phenotypic methods (TI) and by PCR-Restriction Enzyme Analysis (PRA) using the hsp65 gene as target (PRA-hsp65). With 15 of the 32 analysed isolates, results of both methods were concordant, being 8 Mycobacterium avium, 3 M. fortutium, 1 M. kansasii, 1 M. flavescens, 1 M. peregrinum and 1 Nocardia brasiliensis. TI of 12 isolates was inconclusive. Novel PRA-hsp65 patterns were observed with 11 isolates. Medical data were evaluated for inference of clinical relevance of these isolates.
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http://dx.doi.org/10.1590/S1517-838220080002000013DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3768383PMC
April 2008

Mycobacterium kansasii: antibiotic susceptibility and PCR-restriction analysis of clinical isolates.

J Med Microbiol 2005 Oct;54(Pt 10):975-979

Instituto Adolfo Lutz, Sao Paulo, Av. Dr. Arnaldo, 355, Sao Paulo, SP 01246-902, Brazil 2School of Public Health, University of California, Berkeley, CA, USA.

Mycobacterium kansasii is the second most common cause of non-tuberculosis mycobacterial diseases in Sao Paulo, Brazil. An important component of the management of infections caused by this organism is antibiotic susceptibility testing. The objective of this study was to determine the drug susceptibility profiles and genotypes of clinical isolates of M. kansasii obtained from patients with or without an infection that met the American Thoracic Society's case definition criteria of M. kansasii disease. One hundred and sixty-nine clinical isolates of M. kansasii collected between 1993 and 1998 in Sao Paulo, Brazil, were tested consecutively. The isolates were genotyped by PCR restriction-enzyme pattern analysis (PRA). Most of the M. kansasii strains were susceptible to isoniazid, streptomycin, rifabutin, rifampicin, clarithromycin, ethionamide, amikacin, clofazimine and cycloserine, and resistant to ethambutol, ciprofloxacin and doxycycline. Of 169 isolates, 167 belonged to the type I PRA genotype and one each belonged to type II and III genotypes. There was no correlation between PRA subtype and M. kansasii disease according to the American Thoracic Society case definition. Clinical trials may be needed to better correlate MIC values with treatment outcomes to identify appropriate parameters for drug-resistance testing of M. kansasii.
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http://dx.doi.org/10.1099/jmm.0.45965-0DOI Listing
October 2005

Multicenter evaluation of mycobacteria identification by PCR restriction enzyme analysis in laboratories from Latin America and the Caribbean.

J Microbiol Methods 2005 May 25;61(2):193-9. Epub 2004 Dec 25.

Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de São Paulo-Escola Paulista de Medicina, Rua Botucatu, 862 3 degrees andar, São Paulo 04023-062, Brazil.

The identification of mycobacterial species in clinical isolates is essential for making patient care decisions. Polymerase chain reaction (PCR) restriction enzyme analysis (PRA) is a simple and rapid identification method, based on amplification of 441 bp of the hsp65 gene and restriction with BstEII and HaeIII. As a contribution to the validation of PRA, a multicenter study was performed in eight laboratories located in Argentina, Brazil, Colombia, Chile, and Guadeloupe. Each laboratory received 18 coded isolates from the collection of the Institute of Tropical Medicine (Antwerp, Belgium), representing duplicates of nine laboratory strains: Mycobacterium terrae CIPT 140320001, Mycobacterium scrofulaceum CIPT 140220031, Mycobacterium flavescens ATCC 14474, Mycobacterium triviale ATCC 23292, Mycobacterium nonchromogenicum ATCC 19530, Mycobacterium chitae ATCC 19627, Mycobacterium abscessus ATCC 19977, Mycobacterium kansasii ATCC 12478, and Mycobacterium peregrinum ATCC 14467. A detailed protocol including amplification, enzymatic digestion, and gel preparation was provided to each laboratory. Two laboratories identified correctly all 18 (100%) isolates, one identified correctly 17 (94.5%), two identified 14 (77.7%), one identified 11 (61%), and two identified 8 (44.4%) isolates. Errors detected in laboratories with more than 77% accuracy were associated with electrophoresis running conditions and an unspecific amplicon produced by a single strain. Lower accuracy was mainly related to inappropriate use of DNA markers and insufficient training in interpretation of patterns. In conclusion, the PRA method was readily implemented in some Latin American and Caribbean laboratories of mycobacteria, but improvements in critical points, as gel running conditions and training in interpretiation of patterns, are needed in order to improve accuracy. In others, improvement in critical points is still necessary.
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http://dx.doi.org/10.1016/j.mimet.2004.11.015DOI Listing
May 2005

Molecular characterization of Mycobacterium kansasii isolates in the State of São Paulo between 1995-1998.

Mem Inst Oswaldo Cruz 2004 Nov 12;99(7):739-43. Epub 2005 Jan 12.

Setor de Micobactérias, Seção de Bacteriologia, Instituto Adolfo Lutz, São Paulo, Av. Dr. Arnaldo 355, 01246-902, SP, Brazil.

Mycobacterium kansasii is the most common cause of pulmonary nontuberculous mycobacteria infection and classical identification of this pathogen needs a time consuming phenotypic tests. Polymerase chain reaction-restriction fragment length polymorphism analysis (PRA) of the gene enconding for the 65 kDa heat shock (hsp65) protein offers an easy, rapid, and inexpensive procedure to identify and subtype M. kansasii isolates. In the present study, we performed a retrospective analysis of patients who had mycobacteria identified on the basis of phenotypic tests by means of a review of database at Mycobacteria Laboratory of the Instituto Adolfo Lutz in the period 1995-1998. A total of 9381 clinical isolates were analyzed of which 7777 (82.9%) were identified as M. tuberculosis complex and 1604 (17.1%) as nontuberculous mycobacteria. Of the 296 M. kansasii isolates, 189 (63.8%) isolates obtained from 119 patients were viable and were analyzed by PRA-hsp65. Hundred eight two (98.9%) were classified as M. kansasii type I. Two isolates were classified as type II and III and five isolates were characterized as other Mycobacterium species. Clinical isolates of M. kansasii in the state of Sao Paulo was almost exclusively subtype I regardless of HIV status.
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http://dx.doi.org/10.1590/s0074-02762004000700013DOI Listing
November 2004

[Distribution of PRA patterns of clinical isolates of the Mycobacterium avium complex from Spain and South America].

Biomedica 2004 Jun;24 Supp 1:60-4

Departamento de Medicina Preventiva, Facultad de Medicina, Universidad Autónoma, Madrid, España.

Mycobacterium avium complex (MAC) infections are the most frequent systemic infections associated with advanced AIDS. DNA probes for accurate identification of mycobacteria are available but are very expensive in many Latin American settings. Consequently, most Latin American diagnostic laboratories employ inaccurate and outdated tests for mycobacteria identification. Therefore, PCR restriction analysis (PRA) of the hsp65 gene was evaluated for the identification of 163 MAC human isolates originated from Spain and South America. The predominant PRA type in each country was: M. avium type I in Argentina (23/42, 55%) and Brazil (48/72, 67%), M. avium type II in Spain (18/26, 69%) and M. avium type III in Colombia (10/23, 43%). The Colombia frequency is noteworthy, since the PRA type III was quite infrequent in the other three countries. Furthermore, its presence has not been reported outside the Americas. The advantages and disadvantages of PRA in diagnostic mycobacteriology are discussed.
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June 2004
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