Rapid detection of resistance against rifampicin in isolates of from Brazilian patients using a reverse-phase hybridization assay

Oliveira, Martha Maria de; Rocha, Adalgiza da Silva; Oelemann, Maraníbia Cardoso; Gomes, Harisson M.; Fonseca, Leila S.; Werneck-Barreto, A.M.; Valim, Andréia Moura; Rossetti, Maria Lúcia Rosa; Rossau, R.; Mijs, Wouter; Vanderborght, Bart; Suffys, Philip Noël

doi:10.1016/s0167-7012(02)00253-1

Cited by 26 publications

(19 citation statements)

References 25 publications

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“…Other studies have reported significantly lower rates [8,23,24]. This discrepancy may depend on several factors.…”

Section: Discussionmentioning

confidence: 81%

“…To date, heteroresistance has been described for INH, RMP, ethambutol and streptomycin. It is detected using conventional drug susceptibility testing (DST) of several subcultures [5], or by simultaneous detection of wild-type (WT) and mutated sequences using PCR-based techniques, such as restriction fragment length polymorphism [6], sequencing [7] or ''line probe assays'' [8].…”

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confidence: 99%

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Mechanisms of heteroresistance to isoniazid and rifampin of Mycobacterium tuberculosis in Tashkent, Uzbekistan

Hofmann-Thiel¹,

Ingen²,

Feldmann³

et al. 2008

European Respiratory Journal

118

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Heteroresistance of Mycobacterium tuberculosis (MTB) is defined as the coexistence of susceptible and resistant organisms to anti-tuberculosis (TB) drugs in the same patient. Heteroresistance of MTB is considered a preliminary stage to full resistance. To date, no mechanism causing heteroresistance of MTB has been proven.Clinical specimens and cultures from 35 TB patients from Tashkent, Uzbekistan, were analysed using the Genotype MTBDR assay (Hain Lifescience, Nehren, Germany), which is designed to detect genetic mutations associated with resistance to rifampin and isoniazid. Cases of heteroresistance were further subjected to genotyping using mycobacterial interspersed repetitive unit-variable-number tandem repeat typing, spoligotyping and IS6110 fingerprinting.Heteroresistance to rifampin and/or isoniazid was found in seven cases (20%). In five of them, heteroresistance was caused by two different strains and in two by a single strain of the Beijing genotype. The latter cases had a history of relapse of their TB.For the first time, two different mechanisms of heteroresistance in tuberculosis have been proven using a stepwise molecular-biological approach: 1) superinfection with two different strains, which is of interest for clinical infection control practitioners; and 2) splitting of a single strain into susceptible and resistant organisms. The latter mechanism is most likely to be related to poor treatment quality and could serve as a quality marker for tuberculosis therapy programmes in the future.

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“…Other studies have reported significantly lower rates [8,23,24]. This discrepancy may depend on several factors.…”

Section: Discussionmentioning

confidence: 81%

mentioning

confidence: 99%

Mechanisms of heteroresistance to isoniazid and rifampin of Mycobacterium tuberculosis in Tashkent, Uzbekistan

Hofmann-Thiel¹,

Ingen²,

Feldmann³

et al. 2008

European Respiratory Journal

118

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“…A study on RIFr detection of M. tuberculosis from Brazilian patients using reverse-phase hybridization assay was performed by de Oliveira et al (2003). They found that although all sensitive strains had the sensitive genotype, but among resistant isolates, a sensitive genotype was obtained in three due to the absence of mutations in the hot spot region, demonstrating an assay accuracy of 97.6% for detection of drug susceptibility.…”

Section: Discussionmentioning

confidence: 99%

Detection of rifampin resistance patterns in Mycobacterium tuberculosis strains isolated in Iran by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing methods

Isfahani¹,

Tavakoli²,

Salehi³

et al. 2006

Mem. Inst. Oswaldo Cruz

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“…For rpoB, katG, and inhA, the PCR-based sequencing was performed as described by Ramasoota et al (34), de Oliveira et al (35), and Dalla Costa et al (36), while for rpoA and rpoC, the conditions were as described by de Vos et al (37). Sequencing and analysis of rpoA, rpoB, and rpoC genes was performed at Fiocruz-RJ, as described above, while the katG and inhA genes were sequenced and analyzed at the sequencing platform at Centro de Desenvolvimento Científico e Tecnológico (CDCT)-Fundação Estadual de Produção e Pesquisa em Saúde (FEPPS).…”

Section: Methodsmentioning

confidence: 99%

Multidrug-Resistant Mycobacterium tuberculosis of the Latin American Mediterranean Lineage, Wrongly Identified as Mycobacterium pinnipedii (Spoligotype International Type 863 [SIT863]), Causing Active Tuberculosis in South Brazil

et al. 2015

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We recently detected the spoligotype patterns of strains of Mycobacterium pinnipedii, a species of the Mycobacterium tuberculosis complex, in sputum samples from nine cases with pulmonary tuberculosis residing in Porto Alegre, South Brazil. Because this species is rarely encountered in humans, we further characterized these nine isolates by additional genotyping techniques, including 24-locus mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) typing, verification of the loci TbD1, RD9, pks15/1, RD Rio , and fbpC, the insertion of IS6110 at a site specific to the M. tuberculosis Latin American Mediterranean (LAM) lineage, and whole-genome sequencing. The combined analysis of these markers revealed that the isolates are in fact M. tuberculosis and more specifically belong to the LAM genotype. Most of these isolates (n ‫؍‬ 8) were shown to be multidrug resistant (MDR), which prompted us to perform partial sequencing of the rpoA, rpoB, rpoC, katG, and inhA genes. Seven isolates (77.8%) carried the S315T mutation in katG, and one of these (11%) also presented the C(؊17)T single-nucleotide polymorphism (SNP) in inhA. Interestingly, six of the MDR isolates also presented an undescribed insertion of 12 nucleotides (CCA GAA CAA CCC) in codon 516 of rpoB. No putative compensatory mutation was found in either rpoA or rpoC. This is the first report of an M. tuberculosis LAM family strain with a convergent M. pinnipedii spoligotype. These spoligotypes are observed in genotype databases at a modest frequency, highlighting that care must be taken when identifying isolates in the M. tuberculosis complex on the basis of single genetic markers. M. tuberculosis is the predominant cause of human TB worldwide, but M. africanum and M. bovis remain important agents of human disease in certain geographical regions (9). The MTBC species share identical 16S rRNA sequences, and recent studies have improved our knowledge on the genetic diversity, host range, epidemiological aspects, and differences in pathogenicity and virulence among the species of the complex (10, 11). Based on the various genotyping techniques, like spoligotyping (12), restriction fragment length polymorphism (RFLP) (13), mycobacterial interspersed repetitive-unit-variable-number tandem-repeat (MIRU-VNTR) typing (14), and whole-genome sequencing (15), M. tuberculosis strains have been subdivided into lineages and families. The most geographically widespread family worldwide (spoligotyping) is the Latin American Mediterranean (LAM) lineage, which is part of the heterogeneous Euro-American lineage, one of seven M. tuberculosis lineages (16). The LAM family strains are widespread across all five continents, with a marked incidence in

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Rapid detection of resistance against rifampicin in isolates of from Brazilian patients using a reverse-phase hybridization assay

Cited by 26 publications

References 25 publications

Mechanisms of heteroresistance to isoniazid and rifampin of Mycobacterium tuberculosis in Tashkent, Uzbekistan

Mechanisms of heteroresistance to isoniazid and rifampin of Mycobacterium tuberculosis in Tashkent, Uzbekistan

Detection of rifampin resistance patterns in Mycobacterium tuberculosis strains isolated in Iran by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing methods

Multidrug-Resistant Mycobacterium tuberculosis of the Latin American Mediterranean Lineage, Wrongly Identified as Mycobacterium pinnipedii (Spoligotype International Type 863 [SIT863]), Causing Active Tuberculosis in South Brazil

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