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1.
J Clin Microbiol ; 43(12): 5978-82, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16333085

ABSTRACT

Mycobacterium tuberculosis, the etiological agent of tuberculosis, has lost many coding and noncoding regions in its genome during the course of evolution. We performed region-of-difference (RD) analysis using PCR-based genotyping of 131 M. tuberculosis clinical isolates obtained from four different countries, namely, India, Peru, Libya, and Angola. Our studies revealed that RD patterns are often distinct for strains circulating in specific geographical regions and can be used to trace the descent and spread of an isolate from its original reservoir. We describe our findings, which show that no single isolate from the four countries (n = 131) had all the 15 RDs either deleted or retained. Tuberculosis-specific deletion 1 (TbD1) was found to be conserved in 23% of the Indian isolates, indicating their possible ancient origin. RD9 was the most conserved region, RD11 was predominantly deleted, and RD6 was the most variable among the isolates in our collection irrespective of their geographic region. In contrast to earlier reports, our results demonstrate that the deletion of RD1 does not correlate with a decrease in the virulence potential of M. tuberculosis, as Indian isolates (n = 30) examined by us were from diseased individuals and yet had lost the RD1 region. Our results further illustrated that the intactness of the RD5 region may be associated with increased virulence of the organism. This study highlights that the RDs in M. tuberculosis genomes are geographically distributed and specific and may possibly be associated with virulence spectrum.


Subject(s)
Gene Deletion , Genome, Bacterial , Mycobacterium tuberculosis/classification , Polymorphism, Genetic , Tuberculosis/epidemiology , Angola/epidemiology , Bacterial Proteins/genetics , Humans , India/epidemiology , Libya/epidemiology , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Peru/epidemiology , Polymerase Chain Reaction , Tuberculosis/microbiology
2.
J Gastroenterol Hepatol ; 20(10): 1560-6, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16174074

ABSTRACT

BACKGROUND AND AIMS: The genetic composition of the intricate cytotoxin associated gene pathogenicity island (cag PAI) of Helicobacter pylori is known to significantly influence the outcome of the disease. Hence, analysis of complete cag PAI of H. pylori isolated from saliva would be of immense importance in standardizing saliva as a reliable non-invasive diagnostic specimen and also to evaluate the type of H. pylori infection. The aim of the present study was to analyze the genes of cag PAI of H. pylori for their presence and correlating them with the disease status of the patients. METHODS: One hundred and twenty patients (55 duodenal ulcer [DU], 25 gastric ulcer and 40 non-ulcer dyspepsia [NUD]) were investigated for the present study. Eight pairs of oligonucleotide primers (cagA1, cagA2, cagAP1, cagAP2, cagE, cagT, LEC1 and LEC2) of five different loci; cagA, cagA promoter region, cagE which represents cagI region, cagT and LEC representing cagII were used to detect the presence of the cag PAI genes by polymerase chain reaction. RESULTS: The comprehensive analysis of the genes constituting cag PAI showed almost equivalent prevalence of all the genes between both the study groups (ulcer and NUD) included. Little significant difference was found in the percentage distribution in both the clinical groups. cagE and cagT were found in a larger proportion of the ulcer group (92.5% and 96.2%) compared with the NUD group (77.5% and 85%), respectively. CONCLUSION: Saliva could be efficiently used as a non-invasive source for H. pylori and cagT might be an important locus of the cag PAI, thus greatly influencing the disease condition of the subjects.


Subject(s)
Duodenal Ulcer/microbiology , Genomic Islands/genetics , Helicobacter pylori/genetics , Polymerase Chain Reaction , Saliva/microbiology , Stomach Ulcer/microbiology , Adult , Aged , Biopsy , DNA, Bacterial/genetics , Duodenal Ulcer/pathology , Dyspepsia/genetics , Dyspepsia/pathology , Female , Genotype , Humans , Male , Middle Aged , Stomach/microbiology , Stomach/pathology , Stomach Ulcer/pathology , Time Factors
3.
J Clin Microbiol ; 43(8): 4237-41, 2005 Aug.
Article in English | MEDLINE | ID: mdl-16081988

ABSTRACT

The survival and microevolution of Helicobacter pylori strains in the niches of the stomach after eradication therapy have largely been unexplored. We analyzed genomic signatures for two successive isolates obtained 9 years apart from a duodenal ulcer patient who underwent eradication therapy for H. pylori. These isolates were genotyped based on 50 different parameters involving three different fingerprinting approaches and several evolutionarily significant and virulence-associated landmarks in the genome, including nine informative gene loci, the cag pathogenicity island and its right junction, members of the plasticity region cluster, and vacA and iceA alleles. Our observations reveal that the two isolates were derived from the same strain that colonized the patient for almost a decade and were almost identical. Microevolution, however, was observed in the cagA gene and its right junction, the vacA m1 allele, and a member of the plasticity region cluster (JHP926). These results suggest that H. pylori has a great ability to survive and reemerge as a microevolved strain posteradication, thereby hinting at the requirement for follow-up of patients after therapy.


Subject(s)
Duodenal Ulcer/microbiology , Helicobacter Infections/microbiology , Helicobacter pylori/classification , Antigens, Bacterial/genetics , Bacterial Proteins/genetics , Base Sequence , Evolution, Molecular , Genome, Bacterial , Genotype , Helicobacter pylori/genetics , Helicobacter pylori/pathogenicity , Humans , Male , Molecular Sequence Data , Time Factors
4.
BMC Microbiol ; 5: 32, 2005 May 25.
Article in English | MEDLINE | ID: mdl-15916705

ABSTRACT

BACKGROUND: Genomic diversity of H. pylori from many different human populations is largely unknown. We compared genomes of 65 H. pylori strains from Nottingham, England. Molecular analysis was carried out to identify rearrangements within and outside the cag-pathogenicity-island (cag PAI) and DNA sequence divergence in candidate genes. Phylogenetic analysis was carried out based on various high-resolution genotyping techniques. RESULTS: Analyses of virulence genes (cagT, cagE, cagA, vacA, iceA, oipA and babB) revealed that H. pylori strains from England are genetically distinct from strains obtained from other countries. The toxigenic vacA s1m1 genotype was found to be less common and the plasticity region cluster was found to be disrupted in all the isolates. English isolates showed a predominance of iceA1 alleles and a functional proinflammatory oipA gene. The English H. pylori gene pool revealed several Asian/oriental features. This included the predominance of cagA - glr (cagA right junction) motif types III and II (up to 42%), presence of vacA m1c alleles and phylogenetic affinity towards East Asian / Amerindian gene pools based on fluorescent amplified fragment length polymorphism (FAFLP) analysis and glmM sequence analysis. CONCLUSION: Overall, our results demonstrated genetic affinities of H. pylori in England with both European and the Asian gene pools and some distinctive genetic features of virulence genes that may have evolved in this important European population.


Subject(s)
Genomics , Helicobacter Infections/microbiology , Helicobacter pylori/genetics , Helicobacter pylori/isolation & purification , Stomach Ulcer/microbiology , Bacterial Proteins/genetics , England/epidemiology , Genetic Variation , Genome, Bacterial , Helicobacter Infections/epidemiology , Humans , Phylogeny , Stomach Ulcer/epidemiology , Virulence Factors
5.
J Clin Microbiol ; 43(4): 1538-45, 2005 Apr.
Article in English | MEDLINE | ID: mdl-15814963

ABSTRACT

The genomic diversity of Helicobacter pylori from the vast Indian subcontinent is largely unknown. We compared the genomes of 10 H. pylori strains from Ladakh, North India. Molecular analysis was carried out to identify rearrangements within and outside the cag pathogenicity island (cag PAI) and DNA sequence divergence in candidate genes. Analyses of virulence genes (such as the cag PAI as a whole, cagA, vacA, iceA, oipA, babB, and the plasticity cluster) revealed that H. pylori strains from Ladakh are genetically distinct and possibly less virulent than the isolates from East Asian countries, such as China and Japan. Phylogenetic analyses based on the cagA-glr motifs, enterobacterial repetitive intergenic consensus patterns, repetitive extragenic palindromic signatures, the glmM gene mutations, and several genomic markers representing fluorescent amplified fragment length polymorphisms revealed that Ladakhi strains share features of the Indo-European, as well as the East Asian, gene pools. However, the contribution of genetic features from the Indo-European gene pool was more prominent.


Subject(s)
Altitude , Antigens, Bacterial/genetics , Bacterial Proteins/genetics , Desert Climate , Genome, Bacterial , Helicobacter pylori/classification , Helicobacter pylori/pathogenicity , Helicobacter Infections/microbiology , Helicobacter pylori/genetics , Humans , India , Molecular Sequence Data , Phylogeny , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA
6.
J Clin Microbiol ; 42(11): 5302-8, 2004 Nov.
Article in English | MEDLINE | ID: mdl-15528729

ABSTRACT

The cag pathogenicity island (cag-PAI) is one of the major virulence determinants of Helicobacter pylori. The chromosomal integrity of this island or the lack thereof is speculated to play an important role in the progress of the gastroduodenal pathology caused by H. pylori. We determined the integrity of the cag-PAI by using specific flanking and internally anchored PCR primers to know the biogeographical distribution of strains carrying fully integral cag-PAI with proinflammatory behavior in vivo. Genotypes based on eight selected loci were studied in 335 isolates obtained from eight different geographic regions. The cag-PAI appeared to be disrupted in the majority of patient isolates throughout the world. Conservation of cag-PAI was highest in Japanese isolates (57.1%). However, only 18.6% of the Peruvian and 12% of the Indian isolates carried an intact cag-PAI. The integrity of cag-PAI in European and African strains was minimal. All 10 strains from Costa Rica had rearrangements. Overall, a majority of the strains of East Asian ancestry were found to have intact cag-PAI compared to strains of other descent. We also found that the cagE and cagT genes were less often rearranged (18%) than the cagA gene (27%). We attempted to relate cag-PAI rearrangement patterns to disease outcome. Deletion frequencies of cagA, cagE, and cagT genes were higher in benign cases than in isolates from severe ulcers and gastric cancer. Conversely, the cagA promoter and the left end of the cag-PAI were frequently rearranged or deleted in isolates linked to severe pathology. Analysis of the cag-PAI genotypes with a different biogeoclimatic history will contribute to our understanding of the pathogen-host interaction in health and disease.


Subject(s)
Antigens, Bacterial/genetics , Bacterial Proteins/genetics , Gene Deletion , Helicobacter Infections/epidemiology , Helicobacter pylori/pathogenicity , Antigens, Bacterial/metabolism , Asia/epidemiology , Bacterial Proteins/metabolism , Costa Rica/epidemiology , Europe/epidemiology , Genotype , Helicobacter Infections/microbiology , Helicobacter pylori/genetics , Humans , Polymerase Chain Reaction , South Africa/epidemiology , Virulence
7.
J Clin Microbiol ; 42(7): 3240-7, 2004 Jul.
Article in English | MEDLINE | ID: mdl-15243088

ABSTRACT

Tuberculosis continues to be a major killer disease, despite an all-out effort launched against it in the postgenomic era. We describe here the population structure of Mycobacterium tuberculosis strains, as revealed by a chromosome-wide scan of fluorescent amplified fragment length polymorphisms (FAFLPs), for more than 1,100 independent isolates from 11 different countries. The bacterial strains were genotyped based on a total of 136 +/- 1 different FAFLP markers at the genome sequence interface, with details on IS6110 profiles, drug resistance status, clinicopathological observations, and host status integrated into the analysis process. The strains were found to cluster with possible geographic affinities, including the parameters of host species type, IS6110 profile, and drug susceptibility status. Of the five most commonly amplified fragment sets (or amplitypes), type A predominated in strains of mixed origin, deposited in The Netherlands; type B was exclusively observed for Indian isolates; type C was found mainly in strains from Peru and Australia; and types D and E predominated in European strains from France and Italy. The amplitypes were independent of certain large sequence polymorphisms representing two important deletions, TbD1 and Rd9. It appears that M. tuberculosis has a high genomic diversity with a possible geographic evolution. This may have occurred due to specific genomic deletions and synonymous substitutions selected rigorously against host defenses and environmental stresses on an evolutionary timescale. The genotypic data reported here are additionally significant for genotype-phenotype correlations and for determining whether pathogen diversity is a reflection f the host population diversity.


Subject(s)
Mycobacterium tuberculosis/genetics , DNA Transposable Elements , Genome, Bacterial , Genotype , Mycobacterium tuberculosis/classification , Phylogeny , Polymorphism, Genetic
8.
J Clin Microbiol ; 42(6): 2372-8, 2004 Jun.
Article in English | MEDLINE | ID: mdl-15184407

ABSTRACT

Biogeographic partitioning of the genome is typical of the gastric pathogen Helicobacter pylori. Such population-specific evolution could serve as a model for understanding host-pathogen interaction and the impact of genetic drift and recombination on insular populations. With a total of 320 isolates from six geographic regions (Japan, India, England, Spain, Ireland, Africa, and Peru) analyzed by enterobacterial repetitive intergenic consensus (ERIC)-based genotyping, we examined genetic affinities among various H. pylori populations in the world. Several strain-specific and region-specific differences were observed by ERIC-based typing. Polymorphic ERIC patterns indicated that the ERIC sequences are in fact dispersed in the H. pylori chromosome at different locations separated by various distances. Phylogenetic analysis of 61 representative isolates revealed three distinct genetic clusters populated by isolates with shared ERIC types independent of the cag right-junction motif type and vacA allele status. Among the notable genetic relationships were the genotypic similarities between Irish and Japanese and between Peruvian and Japanese isolates. Insular genotypic characteristics of Irish isolates amid genetic similarity to East Asian, as well as North European, strains have been once again proved in this study. Peruvian genotypes were more similar to those of Japanese isolates than to those of Iberian or European isolates. Given the current debate on the origin and age of present-day H. pylori, this is a significant finding that supports the possibility of ancient colonization of Amerindians with East Asian strains. Genotypic data presented here will be additionally helpful in realizing the importance of H. pylori geographical genomics in the development of gastroduodenal pathology.


Subject(s)
DNA Fingerprinting , Enterobacteriaceae/genetics , Helicobacter pylori/classification , Repetitive Sequences, Nucleic Acid , Biological Evolution , Genotype , Helicobacter pylori/genetics , Humans , Multigene Family , Polymerase Chain Reaction , Reproducibility of Results
9.
Bioinformatics ; 20(6): 989-92, 2004 Apr 12.
Article in English | MEDLINE | ID: mdl-14764550

ABSTRACT

AmpliBASE MT is an online databank of high-resolution DNA fingerprints representing fluorescent amplified fragment length polymorphism (FAFLP) profiles or amplitypes developed for the Mycobacterium tuberculosis complex strains from 48 different countries. AmpliBASE MT is based on a relational database management system that is hyperlinked to visualize genotyping results in the form of DNA fingerprint images for individual strains. A flexible search system based on systematic comparisons of fragment sizes in base pairs allows inter-laboratory comparison of FAFLP profiles. Besides this, the database also displays previously published data on IS6110 profiles, spoligotypes, MIRU-VNTRs and large sequence polymorphisms along with the FAFLP records that will give the overall comparisons. Being the first of its kind, AmpliBASE MT is expected to be a very helpful tool in strengthening the concept of 'geographic genomics' and will be very helpful to molecular epidemiologists and those interested in diagnostic development for tuberculosis.


Subject(s)
DNA Fingerprinting/methods , Database Management Systems , Databases, Nucleic Acid , Gene Expression Profiling/methods , Information Storage and Retrieval/methods , Mycobacterium tuberculosis/genetics , User-Computer Interface , Genetic Variation/genetics , Internet , Phenotype , Software
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