ABSTRACT
BACKGROUND: Phylogeographic composition of M. tuberculosis populations reveals associations between lineages and human populations that might have implications for the development of strategies to control the disease. In Latin America, lineage 4 or the Euro-American, is predominant with considerable variations among and within countries. In Colombia, although few studies from specific localities have revealed differences in M. tuberculosis populations, there are still areas of the country where this information is lacking, as is a comparison of Colombian isolates with those from the rest of the world. PRINCIPAL FINDINGS: A total of 414 M. tuberculosis isolates from adult pulmonary tuberculosis cases from three Colombian states were studied. Isolates were genotyped using IS6110-restriction fragment length polymorphism (RFLP), spoligotyping, and 24-locus Mycobacterial interspersed repetitive units variable number tandem repeats (MIRU-VNTRs). SIT42 (LAM9) and SIT62 (H1) represented 53.3% of isolates, followed by 8.21% SIT50 (H3), 5.07% SIT53 (T1), and 3.14% SIT727 (H1). Composite spoligotyping and 24-locus MIRU- VNTR minimum spanning tree analysis suggest a recent expansion of SIT42 and SIT62 evolved originally from SIT53 (T1). The proportion of Haarlem sublineage (44.3%) was significantly higher than that in neighboring countries. Associations were found between M. tuberculosis MDR and SIT45 (H1), as well as HIV-positive serology with SIT727 (H1) and SIT53 (T1). CONCLUSIONS: This study showed the population structure of M. tuberculosis in several regions from Colombia with a dominance of the LAM and Haarlem sublineages, particularly in two major urban settings (Medellín and Cali). Dominant spoligotypes were LAM9 (SIT 42) and Haarlem (SIT62). The proportion of the Haarlem sublineage was higher in Colombia compared to that in neighboring countries, suggesting particular conditions of co-evolution with the corresponding human population that favor the success of this sublineage.
Subject(s)
Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Tuberculosis, Pulmonary/microbiology , Adolescent , Adult , Aged , Aged, 80 and over , Colombia/epidemiology , Female , Genotyping Techniques , Humans , Male , Middle Aged , Phylogeography , Tandem Repeat Sequences/genetics , Young AdultABSTRACT
Homeless people are highly susceptible to tuberculosis. It has been suggested that this population have high rates of mental disorders associated with tuberculosis. We assessed tuberculosis incidence, its transmission patterns and association with socio-demographic factors and mental disorders in Colombian homeless people. Prospective study which socio-demographic characteristics and mental disorders were assessed through interviews. Sputa from patients with respiratory symptoms were processed and clinical isolates analyzed by IS6110-RFLP. Multivariate analysis performed by logistic regression model. From 426 homeless studied, tuberculosis incidence found was 7.9 %. 44 % of isolates were clustering. It was found high risk of having tuberculosis associated with income from drugs trade (OR: 3.40 [95 % CI: 1.28-9.05]), dysthymia (OR: 2.54 [95 % CI: 1.10-5.86]) and receiving food from other homeless (OR: 2.47 [95 % CI: 1.16-5.25]). Tuberculosis incidence and degree of transmission are high in homeless studied. Implementing programs to better control tuberculosis among homeless population must consider socio-demographic factors and mental disorders associated with the disease.
Subject(s)
Ill-Housed Persons/statistics & numerical data , Mental Disorders/epidemiology , Tuberculosis/epidemiology , Adult , Age Factors , Colombia/epidemiology , Female , Ill-Housed Persons/psychology , Humans , Incidence , Male , Middle Aged , Multivariate Analysis , Mycobacterium tuberculosis/isolation & purification , Prospective Studies , Sex Factors , Tuberculosis/transmissionABSTRACT
Tuberculosis is the world's leading cause of death due to a single infectious agent, and efforts aimed at its control require a better understanding of host, environmental, and bacterial factors that govern disease outcome. Growing evidence indicates that certain Mycobacterium tuberculosis strains of distinct phylogeographic lineages elicit unique immunopathological events. However, identifying the genetic basis of these phenotypic peculiarities has proven difficult. Here we report the presence of six large sequence polymorphisms which, together with two single-nucleotide changes previously described by our group, consistently differentiate Haarlem strains from the remaining M. tuberculosis lineages. The six newly found Haarlem-specific genetic events are four deletions, which altogether involve more than 13 kb, and two intragenic insertions of the element IS6110. The absence of the genes involved in these polymorphisms could have an important physiological impact on Haarlem strains, i.e., by affecting key genes, such as Rv1354c and cyp121, which have been recently proposed as plausible drug targets. These lineage-specific polymorphisms can serve as genetic markers for the rapid PCR identification of Haarlem strains, providing a useful tool for strain surveillance and molecular epidemiology studies. Strain variability such as that described here underscores the need for the definition of a core set of essential genes in M. tuberculosis that are ubiquitously present in all circulating lineages, as a requirement in the development of effective antituberculosis drugs and vaccines.
Subject(s)
Antitubercular Agents/pharmacology , DNA, Bacterial/genetics , Mycobacterium tuberculosis/genetics , Polymorphism, Genetic , Tuberculosis Vaccines/immunology , Tuberculosis/microbiology , DNA Transposable Elements , Genetic Markers , Humans , Mutagenesis, Insertional , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/immunology , Polymerase Chain Reaction/methods , Sequence DeletionABSTRACT
Detection of multidrug-resistant tuberculosis (MDR-TB), a frequent cause of treatment failure, takes 2 or more weeks to identify by culture. Rifampicin (RIF) resistance is a hallmark of MDR-TB, and detection of mutations in the rpoB gene of Mycobacterium tuberculosis using molecular beacon probes with real-time quantitative polymerase chain reaction (qPCR) is a novel approach that takes =2 days. However, qPCR identification of resistant isolates, particularly for isolates with mixed RIF-susceptible and RIF-resistant bacteria, is reader dependent and limits its clinical use. The aim of this study was to develop an objective, reader-independent method to define rpoB mutants using beacon qPCR. This would facilitate the transition from a research protocol to the clinical setting, where high-throughput methods with objective interpretation are required. For this, DNAs from 107 M. tuberculosis clinical isolates with known susceptibility to RIF by culture-based methods were obtained from 2 regions where isolates have not previously been subjected to evaluation using molecular beacon qPCR: the Texas-Mexico border and Colombia. Using coded DNA specimens, mutations within an 81-bp hot spot region of rpoB were established by qPCR with 5 beacons spanning this region. Visual and mathematical approaches were used to establish whether the qPCR cycle threshold of the experimental isolate was significantly higher (mutant) compared to a reference wild-type isolate. Visual classification of the beacon qPCR required reader training for strains with a mixture of RIF-susceptible and RIF-resistant bacteria. Only then had the visual interpretation by an experienced reader had 100% sensitivity and 94.6% specificity versus RIF resistance by culture phenotype and 98.1% sensitivity and 100% specificity versus mutations based on DNA sequence. The mathematical approach was 98% sensitive and 94.5% specific versus culture and 96.2% sensitive and 100% specific versus DNA sequence. Our findings indicate the mathematical approach has advantages over the visual reading, in that it uses a Microsoft Excel template to eliminate reader bias or inexperience, and allows objective interpretation from high-throughput analyses even in the presence of a mixture of RIF-resistant and RIF-susceptible isolates without the need for reader training.
Subject(s)
Antitubercular Agents/pharmacology , Bacterial Proteins/genetics , Mutation, Missense , Mycobacterium tuberculosis/drug effects , Polymerase Chain Reaction/methods , Rifampin/pharmacology , Tuberculosis, Multidrug-Resistant/diagnosis , Animals , Colombia , DNA-Directed RNA Polymerases , Humans , Mexico , Microbial Sensitivity Tests/methods , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Sensitivity and Specificity , Texas , Tuberculosis, Multidrug-Resistant/microbiologySubject(s)
Disease Outbreaks , Iatrogenic Disease/epidemiology , Mycobacterium Infections, Nontuberculous/epidemiology , Mycobacterium chelonae/isolation & purification , Skin Diseases, Bacterial/epidemiology , Adolescent , Adult , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , Cluster Analysis , Colombia/epidemiology , DNA Fingerprinting , Female , Genotype , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Mycobacterium Infections, Nontuberculous/microbiology , Random Amplified Polymorphic DNA Technique , Skin Diseases, Bacterial/microbiology , Young AdultABSTRACT
The analysis of the DNA repair genes ogt and ung was carried out in 117 Mycobacterium tuberculosis clinical isolates from Argentina and Colombia in order to explore correlation between mutations in these genes and multi-drug resistance. With the exception of two Beijing family isolates, the rest of the strains harbored either two wild-type or two mutant alleles with identical single nucleotide polymorphisms (SNPs) in each gene (ogt44 and ung501). These ogt44 and ung501 mutations were not associated with multi-drug resistance and occurred simultaneously in circulating Haarlem genotype M. tuberculosis strains. We therefore propose the use of these markers as tools in phylogenetic and epidemiologic studies.
Subject(s)
DNA Repair/genetics , DNA, Bacterial/genetics , Drug Resistance, Multiple, Bacterial/genetics , Mutation , Mycobacterium tuberculosis/genetics , Alleles , Bacterial Proteins/genetics , DNA Mutational Analysis/methods , Genes, Bacterial , Genotype , Humans , Microbial Sensitivity Tests , Mycobacterium tuberculosis/classification , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/isolation & purification , Polymorphism, Single NucleotideABSTRACT
En 157 niños de 2 a 12 años de edad con faringoamigdalitis aguda (FAA), encontramos 85 (54.1 por ciento) con EBH y 66 (42 por ciento) con EBHA. Diecinueve (12.1 por ciento) eran de grupos diferentes al A, siendo el principal el B con 12 casos (7.6 por ciento). Esto ha coincidido con un aislamiento más frecuente de este grupo en orina y con mayor colonización de madres y recién nacidos
Subject(s)
Humans , Child, Preschool , Child , Adolescent , Culture Media , Culture Media/isolation & purification , Diagnosis , Pharyngitis/diagnosis , Signs and Symptoms , Streptococcus/classification , Streptococcus/isolation & purification , Tonsillitis/diagnosis , Enzyme-Linked Immunosorbent AssayABSTRACT
Se revisaron las historias de 107 niños con infección respiratoria aguda (IRA) baja que tenían una IgM positiva para Mycoplasma pneumoniae. La edad más afectada fue la de 2 a 6 años (58 por ciento). El tiempo de evolución antes de la consulta fue de 1 a 180 días, con un promedio de 10.7 días. El motivo de consulta más frecuente fue tos (95.3 por ciento), tos prolongada en el 22.45 por ciento, seguida de fiebre (73.5 por ciento), expectoración y rinitis (32.7 por ciento) respectivamente. Al examen se encontró: Sibilancias (67.3 por ciento), estertores crepitantes (30.8 por ciento) fiebre (37 por ciento), faringitis (15.9 por ciento), otitis (13.1 por ciento) y sinusitis (12 por ciento). El hallazgo radiológico más frecuente fue atrapamiento de aire (21.8 por ciento) derrame pleural abacteriano. La proteína C reactiva fue de < 4 mg por ciento en el 70.8 por ciento. El tratamiento fue a base de macrólidos, principalmente claritromicina (72-6 por ciento), broncodilatadores (40 por ciento) y estos asociados a esteroides en el 25.8 por ciento de los casos
Subject(s)
Humans , Child , Pneumonia, Mycoplasma/complications , Pneumonia, Mycoplasma/diagnosis , Pneumonia, Mycoplasma/epidemiology , Pneumonia, Mycoplasma/etiology , Pneumonia, Mycoplasma/physiopathology , Pneumonia, Mycoplasma/immunology , Pneumonia, Mycoplasma/microbiology , Pneumonia, Mycoplasma/drug therapy , Pneumonia, Mycoplasma , Pneumonia, Mycoplasma/therapy , Respiratory Tract Diseases/diagnosis , Respiratory Tract Diseases/drug therapy , Respiratory Tract Diseases/epidemiology , Respiratory Tract Diseases/etiology , Respiratory Tract Diseases/microbiology , Respiratory Tract Diseases/physiopathologyABSTRACT
Ciento diecinueve niños de uno a 14 años que consultaron por síntomas sugestivos de enfermedad ácido péptica (EAP) fueron estudiados con endoscopia y biopsia. Presentaron gastritis antral 73.1 por ciento, duodenitis 10.08 por ciento, úlcera duodenal 4.2 por ciento, úlcera gástrica 1.6 por ciento y esofagitis 0.8 por ciento. Cincuenta y un pacientes fueron positivos para H.pylori. Tanto el grupo positivo como el negativo tuvieron sintomatología similar, pero la úlcera duodenal se asoció significativamente a H.pylori. Veintinueve (87.8 por ciento) de 33 niños positivos mejoraron clínicamente con diferentes combinaciones de amoxacilina, metronidazol, bismuto y antiácidos y antagonistas H2. Veinticinco (95 por ciento) de 26 negativos mejoraron con antiácidos y antagonistas H2