To understand the genetic diversity and drug resistance status of Mycobocterium tuberculosis （M. tuberculosis） circulating in Xuzhou of China, the spacer-oligonucleotide typing （Spoligotyping） and multi-loci VNTRs...To understand the genetic diversity and drug resistance status of Mycobocterium tuberculosis （M. tuberculosis） circulating in Xuzhou of China, the spacer-oligonucleotide typing （Spoligotyping） and multi-loci VNTRs （variable number tandem repeats） analysis （MLVA） were utilized for the genotyping of the isolates. Drug susceptibility test （DST） was performed by the proportion method on the Lowenstein-Jensen （L-J） medium using isoniazid, rifampicin, ethambutol, and streptomycin. By Spoligotyping, 287 M. tuberculosis isolates were differentiated into 14 clusters. Then with 15-1oci MLVA, these strains could be divided into 32 clusters, 228 genotypes. Of 15 VNTRs, 6 loci had the highly discriminatory powers, 6 loci presented moderate discrimination and 3 loci demonstrated less polymorphism. The DST results showed that 46 strains were resistant to at least one first-line anti-tuberculosis agent. There was a difference in the isoniazid resistance between Beijing and non-Beijing genotype strains. We concluded that the combination of Spoligotyping and 15 VNTR loci as the genotyping in our study was applicable for this region, the drug resistant isolates were identified, and the Beijing family was the most prevalent genotype in the rural counties of Xuzhou.展开更多
Introduction: Tuberculosis is caused by infection with Mycobacterium tuberculosis. Looking at the evolution of the bacterium gene due to mutation is crucial to identify species circulating among patients in an area. W...Introduction: Tuberculosis is caused by infection with Mycobacterium tuberculosis. Looking at the evolution of the bacterium gene due to mutation is crucial to identify species circulating among patients in an area. WHO speculated that tuberculosis is caused by M. tuberculosis (MTB), but identification of the strains of MTB circulating in a particular area is important for the management of MTB and to identify pulmonary infections caused by non-tuberculosis mycobacterium. Contact tracing of drug resistant MTB in circulation in an area is also an important procedure of MTB therapeutic choice. Aim: This study aimed to isolate and identify Mycobacterium species circulating in Bayelsa State, Nigeria. Materials and Methods: A total of 102 sputum samples collected from MTB patients were cultured in Lowenstein Jensen (LJ) solid media. Isolates on LJ media were confirmed using Zeihl Nelseen staining method for AFB and Standard Diagnosis Bioline TB Ag MPT64 Rapid test kit. The 16s rRNA gene amplification, agarose gel electrophoresis, and gene sequencing were conducted. Phylogenic analysis and evolutional distances of the strains are computed using the Juke-cantor method. Result: Out of 102 sputum samples examined 15 (14.7%) had growth of Mycobacterium species (AFB positive). The extracted DNA of MTB amplified on agarose gel electrophoresis aligned horizontally at lanes 1 - 15 showing 16S gene band (1500 bp). Two 2 (2.0%) are non-tuberculosis Mycobacteria species, while 13 (12.7%) were M. tuberculosis. The non-tuberculosis Mycobacterium species isolated are Mycobacteriode abscesses and Mycobacterium kansassii strain FDAARGOS 1534. The tuberculosis strains are Mycobacterium tuberculosis MG003 and R2092 but the predominant strain was MG003. The degree of the genetic evolution of the non-MTB Mycobacterium kansassii strain FDAARGOS 1534 was 75.4%. Conclusion: The two major strains of Mycobacterium tuberculosis (MTB) circulating in Bayelsa State are MTB MG003 and MTB R2092;MTB MG003 was predominant. The non-tuberculosis species are Mycobacteriode abscesses and Mycobacterium kansasii.展开更多
Mycobacterium tuberculosis （Mtb） is a pathogen that is widely distributed geographically and continues to be a major threat to world health. Bacterial virulence factors, nutritional state, host genetic condition an...Mycobacterium tuberculosis （Mtb） is a pathogen that is widely distributed geographically and continues to be a major threat to world health. Bacterial virulence factors, nutritional state, host genetic condition and immune response play an important role in the evolution of the infection. The genetically diverse Mtb strains from different lineages have been shown to induce variable immune system response. The modern and ancient lineages strains induce different cytokines patterns. The immunity to Mtb depends on Th1-cell activity [interferon- γ （IFN- γ ）, interleukin-12 （IL-12） and tumor necrosis factor-α （TNF-α）]. IL-1β directly kills Mtb in murine and human macrophages. IL-6 is a requirement in host resistance to Mtb infection. IFN- γ , TNF-α, IL-12 and IL-17 are participants in Mycobacterium-induced granuloma formation. Other regulating proteins as IL-27 and IL-10 can prevent extensive immunopathology. CXCL 8 enhances the capacity of the neutrophil to kill Mtb . CXCL13 and CCL19 have been identified as participants in the formation of granuloma and control the Mtb infection. Treg cells are increased in patients with active tuberculosis （TB） but decrease with anti-TB treatment. The increment of these cells causes down- regulation of adaptive immune response facilitating the persistence of the bacterial infection. Predominance of Th2 phenotype cytokines increases the severity of TB. The evolution of the Mtb infection will depend of the cytokines network and of the infuence of other factors aforementioned.展开更多
Objective We determined the genetic diversity of Mycobacterium tuberculosis（MTB） in a remote mountainous area of southwest China and evaluated the resolving ability of single nucleotide polymorphism（SNP） genotypin...Objective We determined the genetic diversity of Mycobacterium tuberculosis（MTB） in a remote mountainous area of southwest China and evaluated the resolving ability of single nucleotide polymorphism（SNP） genotyping combined with variable number tandem repeat（VNTR） genotyping for Beijing family strains in association with drug resistance status.Methods Three hundred thirty-one MTB strains were isolated from patients living in mountainous regions of southwest China,and 8-loci SNP,VNTR-15 genotyping assays,and drug susceptibility testing of 9 drugs were performed.Results A total of 183 [55.29%（183/331）] strains were classified into the Beijing family.Of the 183 strains,111（60.66%） were defined as modern Beijing strains.The most predominant modern Beijing sub-lineage and ancient Beijing sub-lineage were Bmyc10 [39.34%（72/183）] and Bmyc25 [20.77%（38/183）],respectively.Of the isolates,19.64%（65/331） were resistant to at least 1 of the 9 anti-TB drugs and 17 [4.98%（17/331）] MTB isolates were multi-drug resistant tuberculosis（MDR-TB）.Two hundred sixty-one isolates showed a clustering rate of 14.18%（37/261） and a discriminatory index of 0.9990.The Beijing lineage exhibited a significantly higher prevalence of MDR-TB,as well as resistance to isoniazid（INH）,rifampin（RIF）,and para-aminosalicylic acid（PAS） when analyzed independently（P = 0.005,P = 0.017,P = 0.014,and P = 0.006 respectively）.The Beijing lineage was not associated with genetic clustering or resistance to any drug.In addition,genetic clustering was not associated with drug resistance.Conclusion MTB strains demonstrate high genetic diversity in remote mountainous areas of southwest China.Beijing strains,especially modern Beijing strains,are predominant in remote mountainous area of China.The combination of 8-loci SNPs and VNTR-15 genotyping is a useful tool to study the molecular epidemiology of MTB strains in this area.展开更多
Several related substances were detected at trace level in (2R)-2,3-dihydro-2-methyl-6-nitro-2-[[4-[4-[4-(trifluoromethoxy)phenoxy]-1-piperidinyl] phenoxy] methyl]imidazo[2, 1-b]oxazole drug substance by a newly devel...Several related substances were detected at trace level in (2R)-2,3-dihydro-2-methyl-6-nitro-2-[[4-[4-[4-(trifluoromethoxy)phenoxy]-1-piperidinyl] phenoxy] methyl]imidazo[2, 1-b]oxazole drug substance by a newly developed high-performance liquid chromatography method. All related substances were characterized rapidly but some impurities were found to be intermediates. Proposed structures were further confirmed by characterization using NMR, FT-IR, and HRMS techniques. Based on the spectroscopic data;unknown related sub-stances were characterized as 1-(Methylsulfonyl)-4-[4-(trifluoromethoxy) phenoxy]piperidine;4-{4-[4-(Tri-fluoromethoxy)-phenoxy]piperidin-1-yl}phenol and 4-{4-[4-(trifluoromethoxy)phenoxy]piperidin-1-yl}phenyl methane sulfonate;4-Bromophenyl methane sulfonate, Ethyl 3,6-dihydro-1(2H)-pyridine carboxylate, (2S)-3-(4-Bromophenoxy)-2-hydroxy-2-methylpropyl methane sulfonate, (2S)-3-(4-Bromophenoxy)-2-methylpropane-1,2-diyldimethane-sulfonate, (2S)-2-Methyl-3-(4-{4-[4-(trifluoromethoxy) phenoxy]-piperidin-1-yl} phenoxy)-propane-1,2-diyldimethane sulfonate, (S)-3-(4-Bromophenoxy)-2-methyl-propane-1,2-diol and corresponding Enantiomer, (2R)-2-[(4-Bromo-phenoxy)methyl]-2-methyloxirane and (2R)-2-[(4-bromophenoxy)methyl]-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b][1,3]oxazole. A possible mechanism for the formation of these related substances is also proposed.展开更多
基金funded by the projects 2013ZX10003002-001 of Chinese National Key Program of Mega Infectious Disease of the National 12th Five-Year Planthe Science and Technology Innovation Team Support project CX201412 of Changzhi Medical College
文摘To understand the genetic diversity and drug resistance status of Mycobocterium tuberculosis （M. tuberculosis） circulating in Xuzhou of China, the spacer-oligonucleotide typing （Spoligotyping） and multi-loci VNTRs （variable number tandem repeats） analysis （MLVA） were utilized for the genotyping of the isolates. Drug susceptibility test （DST） was performed by the proportion method on the Lowenstein-Jensen （L-J） medium using isoniazid, rifampicin, ethambutol, and streptomycin. By Spoligotyping, 287 M. tuberculosis isolates were differentiated into 14 clusters. Then with 15-1oci MLVA, these strains could be divided into 32 clusters, 228 genotypes. Of 15 VNTRs, 6 loci had the highly discriminatory powers, 6 loci presented moderate discrimination and 3 loci demonstrated less polymorphism. The DST results showed that 46 strains were resistant to at least one first-line anti-tuberculosis agent. There was a difference in the isoniazid resistance between Beijing and non-Beijing genotype strains. We concluded that the combination of Spoligotyping and 15 VNTR loci as the genotyping in our study was applicable for this region, the drug resistant isolates were identified, and the Beijing family was the most prevalent genotype in the rural counties of Xuzhou.
文摘Introduction: Tuberculosis is caused by infection with Mycobacterium tuberculosis. Looking at the evolution of the bacterium gene due to mutation is crucial to identify species circulating among patients in an area. WHO speculated that tuberculosis is caused by M. tuberculosis (MTB), but identification of the strains of MTB circulating in a particular area is important for the management of MTB and to identify pulmonary infections caused by non-tuberculosis mycobacterium. Contact tracing of drug resistant MTB in circulation in an area is also an important procedure of MTB therapeutic choice. Aim: This study aimed to isolate and identify Mycobacterium species circulating in Bayelsa State, Nigeria. Materials and Methods: A total of 102 sputum samples collected from MTB patients were cultured in Lowenstein Jensen (LJ) solid media. Isolates on LJ media were confirmed using Zeihl Nelseen staining method for AFB and Standard Diagnosis Bioline TB Ag MPT64 Rapid test kit. The 16s rRNA gene amplification, agarose gel electrophoresis, and gene sequencing were conducted. Phylogenic analysis and evolutional distances of the strains are computed using the Juke-cantor method. Result: Out of 102 sputum samples examined 15 (14.7%) had growth of Mycobacterium species (AFB positive). The extracted DNA of MTB amplified on agarose gel electrophoresis aligned horizontally at lanes 1 - 15 showing 16S gene band (1500 bp). Two 2 (2.0%) are non-tuberculosis Mycobacteria species, while 13 (12.7%) were M. tuberculosis. The non-tuberculosis Mycobacterium species isolated are Mycobacteriode abscesses and Mycobacterium kansassii strain FDAARGOS 1534. The tuberculosis strains are Mycobacterium tuberculosis MG003 and R2092 but the predominant strain was MG003. The degree of the genetic evolution of the non-MTB Mycobacterium kansassii strain FDAARGOS 1534 was 75.4%. Conclusion: The two major strains of Mycobacterium tuberculosis (MTB) circulating in Bayelsa State are MTB MG003 and MTB R2092;MTB MG003 was predominant. The non-tuberculosis species are Mycobacteriode abscesses and Mycobacterium kansasii.
基金Supported by Institute of Biological Research,Faculty of Medicine,University of Zulia,Maracaibo,Venezuela
文摘Mycobacterium tuberculosis （Mtb） is a pathogen that is widely distributed geographically and continues to be a major threat to world health. Bacterial virulence factors, nutritional state, host genetic condition and immune response play an important role in the evolution of the infection. The genetically diverse Mtb strains from different lineages have been shown to induce variable immune system response. The modern and ancient lineages strains induce different cytokines patterns. The immunity to Mtb depends on Th1-cell activity [interferon- γ （IFN- γ ）, interleukin-12 （IL-12） and tumor necrosis factor-α （TNF-α）]. IL-1β directly kills Mtb in murine and human macrophages. IL-6 is a requirement in host resistance to Mtb infection. IFN- γ , TNF-α, IL-12 and IL-17 are participants in Mycobacterium-induced granuloma formation. Other regulating proteins as IL-27 and IL-10 can prevent extensive immunopathology. CXCL 8 enhances the capacity of the neutrophil to kill Mtb . CXCL13 and CCL19 have been identified as participants in the formation of granuloma and control the Mtb infection. Treg cells are increased in patients with active tuberculosis （TB） but decrease with anti-TB treatment. The increment of these cells causes down- regulation of adaptive immune response facilitating the persistence of the bacterial infection. Predominance of Th2 phenotype cytokines increases the severity of TB. The evolution of the Mtb infection will depend of the cytokines network and of the infuence of other factors aforementioned.
基金supported by the National Science and Technology Major Project 2014ZX100030002The National Basic Research Program of China 2014CB744403
文摘Objective We determined the genetic diversity of Mycobacterium tuberculosis（MTB） in a remote mountainous area of southwest China and evaluated the resolving ability of single nucleotide polymorphism（SNP） genotyping combined with variable number tandem repeat（VNTR） genotyping for Beijing family strains in association with drug resistance status.Methods Three hundred thirty-one MTB strains were isolated from patients living in mountainous regions of southwest China,and 8-loci SNP,VNTR-15 genotyping assays,and drug susceptibility testing of 9 drugs were performed.Results A total of 183 [55.29%（183/331）] strains were classified into the Beijing family.Of the 183 strains,111（60.66%） were defined as modern Beijing strains.The most predominant modern Beijing sub-lineage and ancient Beijing sub-lineage were Bmyc10 [39.34%（72/183）] and Bmyc25 [20.77%（38/183）],respectively.Of the isolates,19.64%（65/331） were resistant to at least 1 of the 9 anti-TB drugs and 17 [4.98%（17/331）] MTB isolates were multi-drug resistant tuberculosis（MDR-TB）.Two hundred sixty-one isolates showed a clustering rate of 14.18%（37/261） and a discriminatory index of 0.9990.The Beijing lineage exhibited a significantly higher prevalence of MDR-TB,as well as resistance to isoniazid（INH）,rifampin（RIF）,and para-aminosalicylic acid（PAS） when analyzed independently（P = 0.005,P = 0.017,P = 0.014,and P = 0.006 respectively）.The Beijing lineage was not associated with genetic clustering or resistance to any drug.In addition,genetic clustering was not associated with drug resistance.Conclusion MTB strains demonstrate high genetic diversity in remote mountainous areas of southwest China.Beijing strains,especially modern Beijing strains,are predominant in remote mountainous area of China.The combination of 8-loci SNPs and VNTR-15 genotyping is a useful tool to study the molecular epidemiology of MTB strains in this area.
文摘Several related substances were detected at trace level in (2R)-2,3-dihydro-2-methyl-6-nitro-2-[[4-[4-[4-(trifluoromethoxy)phenoxy]-1-piperidinyl] phenoxy] methyl]imidazo[2, 1-b]oxazole drug substance by a newly developed high-performance liquid chromatography method. All related substances were characterized rapidly but some impurities were found to be intermediates. Proposed structures were further confirmed by characterization using NMR, FT-IR, and HRMS techniques. Based on the spectroscopic data;unknown related sub-stances were characterized as 1-(Methylsulfonyl)-4-[4-(trifluoromethoxy) phenoxy]piperidine;4-{4-[4-(Tri-fluoromethoxy)-phenoxy]piperidin-1-yl}phenol and 4-{4-[4-(trifluoromethoxy)phenoxy]piperidin-1-yl}phenyl methane sulfonate;4-Bromophenyl methane sulfonate, Ethyl 3,6-dihydro-1(2H)-pyridine carboxylate, (2S)-3-(4-Bromophenoxy)-2-hydroxy-2-methylpropyl methane sulfonate, (2S)-3-(4-Bromophenoxy)-2-methylpropane-1,2-diyldimethane-sulfonate, (2S)-2-Methyl-3-(4-{4-[4-(trifluoromethoxy) phenoxy]-piperidin-1-yl} phenoxy)-propane-1,2-diyldimethane sulfonate, (S)-3-(4-Bromophenoxy)-2-methyl-propane-1,2-diol and corresponding Enantiomer, (2R)-2-[(4-Bromo-phenoxy)methyl]-2-methyloxirane and (2R)-2-[(4-bromophenoxy)methyl]-2-methyl-6-nitro-2,3-dihydroimidazo[2,1-b][1,3]oxazole. A possible mechanism for the formation of these related substances is also proposed.
