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1.
J Natl Cancer Inst ; 99(14): 1086-94, 2007 Jul 18.
Article in English | MEDLINE | ID: mdl-17623796

ABSTRACT

BACKGROUND: Kaposi sarcoma (KS) is a complex tumor of uncertain clonality. Studying the viral clonality of the human herpesvirus 8 (HHV-8) in KS to determine clonality of the tumors, a strategy that has been used previously with Epstein-Barr virus and its associated tumors, may elucidate whether multicentric (disseminated) KS lesions correspond to metastatic lesions or to expansions of independent clones. METHODS: A series of 139 KS biopsies (from skin, lymph node, or tonsil) was obtained from 98 patients, with 59 biopsies from 18 patients with disseminated multicentric KS skin lesions. The degree of spindle cell infiltration in biopsies was established by direct observation of hematoxylin-eosin-stained sections, and HHV-8 viral load was quantified by real-time polymerase chain reaction. To determine cellular clonality, the size heterogeneity of the HHV-8-fused terminal repeat (TR) region was determined by probing of electrophoresed restricted genomic DNA from KS biopsies for the HHV-8 TR sequence. RESULTS: HHV-8 clonality analysis was performed on the 62 samples for which sufficient DNA was obtained. Most samples corresponded to histologically nodular lesions with high spindle cell infiltration and high viral load. A clonal HHV-8 pattern was determined for 59 samples; 11 were found to be monoclonal and 48 to be oligoclonal. The informative samples that were from disseminated KS skin lesions (n = 26, from six patients) were either monoclonal or oligoclonal, and the size of HHV-8 episomes varied between these samples. CONCLUSION: Although some tumor KS lesions were monoclonal expansions of HHV-8-infected spindle cells, most advanced lesions were oligoclonal proliferations. Furthermore, individual KS disseminated tumor skin lesions were found to represent distinct expansions of HHV-8-infected spindle cells. Thus, our results suggest that KS lesions, especially in patients with advanced skin tumors, are reactive proliferations rather than true malignancies with metastatic dissemination.


Subject(s)
Herpesvirus 8, Human/genetics , Sarcoma, Kaposi/pathology , Sarcoma, Kaposi/virology , Skin/pathology , Adolescent , Adult , Aged , Aged, 80 and over , Child , Clone Cells/pathology , Clone Cells/virology , DNA, Viral/analysis , DNA, Viral/genetics , Female , Herpesvirus 8, Human/isolation & purification , Humans , Male , Middle Aged , Viral Load
2.
BMC Microbiol ; 7: 39, 2007 May 16.
Article in English | MEDLINE | ID: mdl-17506895

ABSTRACT

BACKGROUND: Previous studies have suggested that variations in DNA repair genes of W-Beijing strains may have led to transient mutator phenotypes which in turn may have contributed to host adaptation of this strain family. Single nucleotide polymorphism (SNP) in the DNA repair gene mutT1 was identified in MDR-prone strains from the Central African Republic. A Mycobacteriumtuberculosis H37Rv mutant inactivated in two DNA repair genes, namely ada/alkA and ogt, was shown to display a hypermutator phenotype. We then looked for polymorphisms in these genes in Central African Republic strains (CAR). RESULTS: In this study, 55 MDR and 194 non-MDR strains were analyzed. Variations in DNA repair genes ada/alkA and ogt were identified. Among them, by comparison to M. tuberculosis published sequences, we found a non-sense variation in ada/alkA gene which was also observed in M. bovis AF2122 strain. SNPs that are present in the adjacent regions to the amber variation are different in M. bovis and in M. tuberculosis strain. CONCLUSION: An Amber codon was found in the ada/alkA locus of clustered M. tuberculosis isolates and in M. bovis strain AF2122. This is likely due to convergent evolution because SNP differences between strains are incompatible with horizontal transfer of an entire gene. This suggests that such a variation may confer a selective advantage and be implicated in hypermutator phenotype expression, which in turn contributes to adaptation to environmental changes.


Subject(s)
Bacterial Proteins/genetics , Codon, Nonsense/genetics , Evolution, Molecular , Mycobacterium bovis/genetics , Mycobacterium tuberculosis/genetics , Polymorphism, Single Nucleotide , Central African Republic , DNA Repair Enzymes/genetics , Humans , Mycobacterium bovis/isolation & purification , Mycobacterium tuberculosis/isolation & purification , Phenotype , Tuberculosis/microbiology
3.
Emerg Infect Dis ; 12(9): 1454-6, 2006 Sep.
Article in English | MEDLINE | ID: mdl-17073103

ABSTRACT

We investigated multidrug-resistant (MDR) Mycobacterium tuberculosis strains in Bangui, Central African Republic. We found 39.6% with the same spoligotype and synonymous single nucleotide polymorphism in the mutT1 gene. However, strains had different rpoB mutations responsible for rifampin resistance. MDR strains in Bangui may emerge preferentially from a single, MDR-prone family.


Subject(s)
Mycobacterium tuberculosis , Tuberculosis, Multidrug-Resistant/epidemiology , Tuberculosis, Multidrug-Resistant/microbiology , Antitubercular Agents/pharmacology , Bacterial Proteins/genetics , Central African Republic/epidemiology , DNA Repair , DNA-Directed RNA Polymerases , Humans , Mutation , Mycobacterium tuberculosis/classification , Mycobacterium tuberculosis/drug effects , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Oligonucleotides/analysis , Polymorphism, Single Nucleotide
4.
Clin Vaccine Immunol ; 13(6): 702-3, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16760330

ABSTRACT

We assessed the performance of a serological test for tuberculosis (SDHO Laboratories Inc., Canada) in our setting. Among 68 of 99 suspected pulmonary tuberculosis patients who were scored as having tuberculosis on the basis of Mycobacterium tuberculosis-positive culture, the sensitivity of the serological test was lower than that of sputum smear microscopic examination (20.6% versus 80.9%, respectively; P < 0.000001).


Subject(s)
Serologic Tests/methods , Tuberculosis, Pulmonary/blood , Tuberculosis, Pulmonary/diagnosis , Adolescent , Adult , Aged , Central African Republic/epidemiology , Female , Humans , Male , Middle Aged , Tuberculosis, Pulmonary/epidemiology
5.
J Clin Microbiol ; 43(9): 4840-3, 2005 Sep.
Article in English | MEDLINE | ID: mdl-16145154

ABSTRACT

Epidemic Kaposi's sarcoma (KS) is one of the most frequent types of cancer in several African countries; however, very few data are available on human herpesvirus 8 (HHV-8) markers in KS patients from Central Africa. In a series of 36 AIDS-KS cases from Central African Republic, we showed, using a real-time PCR quantitative assay, the high frequency (82%) of detectable HHV-8 DNA in peripheral blood mononuclear cells (PBMCs). We also found that the level of antibodies directed against lytic or latent HHV-8 antigens is not correlated to the amount of HHV-8 viral load in the PBMCs, and finally, we demonstrated a much higher viral load in tumoral skin lesions (6.07 log copies/mug DNA) than in unaffected skin (2.93 log copies/mug DNA) or in PBMCs (2.55 log copies/mug DNA).


Subject(s)
AIDS-Related Opportunistic Infections/virology , Antibodies, Viral/blood , DNA, Viral/blood , Herpesvirus 8, Human/isolation & purification , Sarcoma, Kaposi/virology , Skin Neoplasms/virology , Viral Load , AIDS-Related Opportunistic Infections/immunology , Adolescent , Adult , Antigens, Viral/immunology , Central African Republic , Child , DNA, Viral/analysis , Female , HIV Infections , Herpesvirus 8, Human/genetics , Herpesvirus 8, Human/immunology , Humans , Leukocytes, Mononuclear/virology , Male , Middle Aged , Polymerase Chain Reaction , Sarcoma, Kaposi/immunology , Skin/virology
6.
Acta Trop ; 89(1): 67-72, 2003 Dec.
Article in English | MEDLINE | ID: mdl-14636984

ABSTRACT

To determine which pathogens are responsible for bloodstream infections in Bangui and to which antibiotics these pathogens are resistant, we conducted a prospective study of the bacteria isolated from the blood of febrile patients hospitalized in the department of medicine of the Bangui Community Hospital after the failure of antimalarial treatment. One hundred and thirty-one patients were included in this study. Bacteria were identified in 49 blood cultures (37.4%). Eleven different species were identified. Bacteremia was more frequent in HIV-positive patients than in HIV-negative patients. Salmonella typhimurium, Mycobacterium tuberculosis and Streptococcus pneumoniae were the most frequently isolated pathogens. Eighty percent of enterobacteria were resistant to amoxicillin and 85% to trimethoprim-sulfamethoxazole. Ciprofloxacin and ceftriaxone were the most efficient antibiotics for the enterobacteria, but chloramphenicol and gentamicin were efficient in most cases. Some strains of S. pneumoniae displayed reduced susceptibility to penicillin G, but all strains were susceptible to erythromycin.


Subject(s)
Anti-Bacterial Agents/therapeutic use , Bacteremia/microbiology , Bacteria/isolation & purification , Adolescent , Adult , Bacteremia/drug therapy , Bacteremia/epidemiology , Bacteria/drug effects , Bacteria/pathogenicity , Central African Republic/epidemiology , Drug Resistance, Bacterial , Enzyme-Linked Immunosorbent Assay , Female , HIV Infections/complications , Hospitalization , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Prospective Studies
7.
J Clin Microbiol ; 41(5): 1963-70, 2003 May.
Article in English | MEDLINE | ID: mdl-12734235

ABSTRACT

The present update on the global distribution of Mycobacterium tuberculosis complex spoligotypes provides both the octal and binary descriptions of the spoligotypes for M. tuberculosis complex, including Mycobacterium bovis, from >90 countries (13,008 patterns grouped into 813 shared types containing 11,708 isolates and 1,300 orphan patterns). A number of potential indices were developed to summarize the information on the biogeographical specificity of a given shared type, as well as its geographical spreading (matching code and spreading index, respectively). To facilitate the analysis of hundreds of spoligotypes each made up of a binary succession of 43 bits of information, a number of major and minor visual rules were also defined. A total of six major rules (A to F) with the precise description of the extra missing spacers (minor rules) were used to define 36 major clades (or families) of M. tuberculosis. Some major clades identified were the East African-Indian (EAI) clade, the Beijing clade, the Haarlem clade, the Latin American and Mediterranean (LAM) clade, the Central Asian (CAS) clade, a European clade of IS6110 low banders (X; highly prevalent in the United States and United Kingdom), and a widespread yet poorly defined clade (T). When the visual rules defined above were used for an automated labeling of the 813 shared types to define nine superfamilies of strains (Mycobacterium africanum, Beijing, M. bovis, EAI, CAS, T, Haarlem, X, and LAM), 96.9% of the shared types received a label, showing the potential for automated labeling of M. tuberculosis families in well-defined phylogeographical families. Intercontinental matches of shared types among eight continents and subcontinents (Africa, North America, Central America, South America, Europe, the Middle East and Central Asia, and the Far East) are analyzed and discussed.


Subject(s)
Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Bacterial Typing Techniques , DNA, Bacterial/genetics , DNA, Intergenic/genetics , Databases, Nucleic Acid , Humans , Molecular Epidemiology , Mycobacterium bovis/classification , Mycobacterium bovis/genetics , Mycobacterium bovis/isolation & purification , Mycobacterium tuberculosis/classification , Tuberculosis/epidemiology , Tuberculosis/microbiology
8.
Emerg Infect Dis ; 8(11): 1347-9, 2002 Nov.
Article in English | MEDLINE | ID: mdl-12453368

ABSTRACT

We present a short summary of recent observations on the global distribution of the major clades of the Mycobacterium tuberculosis complex, the causative agent of tuberculosis. This global distribution was defined by data-mining of an international spoligotyping database, SpolDB3. This database contains 11708 patterns from as many clinical isolates originating from more than 90 countries. The 11708 spoligotypes were clustered into 813 shared types. A total of 1300 orphan patterns (clinical isolates showing a unique spoligotype) were also detected.


Subject(s)
Mycobacterium tuberculosis/classification , Mycobacterium tuberculosis/isolation & purification , Tuberculosis/microbiology , Databases, Factual , Phylogeny , Tuberculosis/epidemiology
9.
Microbiology (Reading) ; 145 ( Pt 7): 1695-1701, 1999 Jul.
Article in English | MEDLINE | ID: mdl-10439408

ABSTRACT

It has previously been shown that the PAN promoter from Mycobacterium paratuberculosis can be used as a DNA probe to identify an RFLP between wild-type Mycobacterium bovis and the vaccine strain Mycobacterium bovis BCG. To investigate the genetic basis of this phenomenon, DNA fragments from a New Zealand M. bovis cattle strain and M. bovis BCG Pasteur, containing the PAN-binding region, were isolated from gene libraries, sequenced and characterized. Sequence analysis and comparison with database sequences showed that the PAN region in M. bovis, M. bovis BCG and Mycobacterium tuberculosis is identical and shares 70% similarity to the PAN sequence from M. paratuberculosis. The Shine-Dalgarno sequence and the -10 and -35 promoter regions are conserved between the different species. Analysis of the flanking sequences of the PAN region revealed that less than 1 kb downstream of PAN is a 2405 bp fragment that is present in M. bovis BCG but absent in the M. bovis wild-type strain. The distribution of the 2405 bp fragment in members of the M. tuberculosis complex was investigated and found to be present in 70 out of 70 M. tuberculosis strains, and 7 out of 7 M. bovis BCG daughter strains, 2 out of 2 Mycobacterium africanum strains, 2 out of 2 Mycobacterium microti strains and 7 out of 25 M. bovis strains. This is the first report of a genetic region of M. bovis BCG that is not universally present in M. bovis strains. The fragment does not appear to be present in any mycobacterial species outside the M. tuberculosis complex. It does not possess any characteristics of known transposable elements and the flanking sequences do not have any obvious features to suggest a deletion mechanism. The genetic location of this region is close to the 3' end of the RD1 region of M. bovis and M. tuberculosis. The polymorphic nature of this locus in M. bovis will provide an additional genetic marker for strain differentiation.


Subject(s)
Mycobacterium bovis/genetics , Mycobacterium tuberculosis/genetics , Polymorphism, Restriction Fragment Length , Promoter Regions, Genetic , Amino Acid Sequence , Animals , Base Sequence , Blotting, Southern , Cattle , DNA Probes , DNA, Bacterial/genetics , Humans , Molecular Sequence Data , Mycobacterium bovis/classification , Mycobacterium tuberculosis/classification , Nontuberculous Mycobacteria/classification , Nontuberculous Mycobacteria/genetics , Polymerase Chain Reaction , Sequence Analysis, DNA
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