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ABSTRACT: Mycoplasma pneumoniae (M. pneumoniae) is one of the most important pathogens that cause respiratory tract infection in children and adults. In this study, we describe a rapid and sensitive colorimetric loop mediated isothermal amplification (LAMP) method to detect M. pneumoniae. The specificity and sensitivity of this assay were detected with 21 common respiratory pathogens and 39 M. pneumoniae DNA. The sensitivity of LAMP was 100% among 39 M. pneumoniae isolates and the specificity was 100% among 9 members of other Mycoplasma and 12 common respiratory pathogens. The lowest detectable limit (LDL) of this assay was 102 copies, which detected by a series of standard M. pneumoniae DNA. To evaluate the clinical applicability of the LAMP assay, a total of 80 clinical samples were examined by conventional PCR, real-time PCR and the LAMP assays, respectively. The positive rates were 15.0%, 32.5% and 26.3%, respectively. This colorimetric LAMP assay demonstrated a high level of sensitivity comparable with that of conventional PCR for the detection of M. pneumoniae. It is a valuable method for simple, cost-effective and rapid detection of M. pneumoniae in the rural areas and basic clinical of China.
Acta Microbiologica et Immunologica Hungarica 03/2013; 60(1):1-9. · 0.79 Impact Factor
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Fei Zhao,
Gang Liu,
Jiang Wu,
Bin Cao,
Xiaoxia Tao, Lihua He,
Fanliang Meng,
Liang Zhu,
Min Lv,
Yudong Yin,
Jianzhong Zhang
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ABSTRACT: Macrolide resistance rates of Mycoplasma pneumoniae in Beijing population were as high as 68.9%, 90.0%, 98.4%, 95.4% and 97.0% from 2008 to 2012, respectively. Common macrolide resistant mobile genetic elements were not detected with any isolate. These macrolide resistant isolates came from multiple clones rather than the same clone. No massive aggregation of a particular clone was found in specific period.
Antimicrobial Agents and Chemotherapy 12/2012; · 4.84 Impact Factor
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Fei Zhao,
Gang Liu,
Bin Cao,
Jiang Wu,
Yixin Gu, Lihua He,
Fanliang Meng,
Liang Zhu,
Yudong Yin,
Min Lv,
Jianzhong Zhang
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ABSTRACT: 201 Mycoplasma pneumoniae clinical isolates isolated from Beijing in 2008 to 2011 were clustered into 16 Multiple-locus Variable number tandem repeat Analysis (MLVA) types, of which 6 new MLVA types have never been previously reported. Type 1 isolates based on p1 gene genotyping were mainly MLVA types E, J, P, U and X. There was no correlation between macrolide-resistant Mycoplasma pneumoniae and their MLVA type.
Journal of clinical microbiology 12/2012; · 4.16 Impact Factor
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ABSTRACT: Helicobacter pylori is a bacterial pathogen which can lead to several human gastric diseases. Here we describe the genome sequences of three strains isolated from atrophic gastritis and gastric ulcers patients in China. The data will permit genomic characterization of traits that may contribute to various gastric diseases.
Journal of bacteriology 11/2012; 194(22):6314-5. · 3.94 Impact Factor
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ABSTRACT: In this study, a whole-genome CombiMatrix Custom oligonucleotide tiling microarray with 90,000 probes covering six sequenced Helicobacter pylori (H. pylori) genomes was designed. This microarray was used to compare the genomic profiles of eight unsequenced strains isolated from patients with different gastroduodenal diseases in Heilongjiang province of China. Since significant genomic variation was found among these strains, an additional 76 H. pylori strains associated with different clinical outcomes were isolated from various provinces of China. These strains were tested by polymerase chain reaction to demonstrate this distinction. We identified several highly variable regions in strains associated with gastritis, gastric ulceration, and gastric cancer. These regions are associated with genes involved in the bacterial type I, type II, and type III R-M systems. They were also associated with the virB gene, which lies on the well-studied cag pathogenic island. While previous studies have reported on the diverse genetic characterization of this pathogenic island, in this study, we find that it is conserved in all strains tested by microarray. Moreover, a number of genes involved in the type IV secretion system, which is related to horizontal DNA transfer between H. pylori strains, were identified in the comparative analysis of the strain-specific genes. These findings may provide insight into new biomarkers for the prediction of gastric diseases.
PLoS ONE 01/2012; 7(6):e38528. · 4.09 Impact Factor
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ABSTRACT: The antimicrobials resistance of Helicobacter pylori (H. pylori) was able to sharply decline the eradication rate of H. pylori both in adults and children, but there are limited studies about the primary antibiotic resistance and the related gene mutations, specifically in China.
The primary resistance to 9 antibiotics of 73 H. pylori strains isolated from gastric biopsies of children recruited at Beijing Children's Hospital was assessed, and the mutations in 23S rRNA gene of 65 macrolide-resistant strains and in gyrA and gyrB of 12 quinolone-resistant strains were investigated.
The resistance rate to clarithromycin, azithromycin, metronidazole, levofloxacin, moxifloxacin, and rifampicin was 84.9%, 87.7%, 61.6%, 13.7%, 15.1%, and 6.8%, respectively. No resistance to amoxicillin, gentamicin, and tetracycline was observed. Dual, triple, and quadruple antibacterial resistant percentage was 46.6% (34/73), 15.1% (11/73), and 2.7% (2/73), respectively. The gene mutation rate of A2142C, A2142G, and A2143G in 23S rRNA gene was 1.5% (1/65), 6.2% (4/65), and 84.6% (55/65), respectively. The detection rate of mutations of Asn87, Asp91, and Met191 in GyrA was 41.7% (5/12), 25% (3/12), and 25% (3/12), respectively.
The high prevalence of primary antibiotic resistance was out of expectation in H. pylori strains isolated from the children in Beijing. Antibiotic susceptibility should be made clear before the antibiotic was used in the anti-H. pylori therapy in this population. The A2143G was the most populated mutation in macrolide-resistant strains, and Asn87 and Asp91 of GyrA were the most common mutation points in quinolone resistance strains.
Helicobacter 10/2011; 16(5):356-62. · 3.15 Impact Factor
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ABSTRACT: The p1 genes of 60 Mycoplasma pneumoniae clinical isolates were sequenced and compared to previously reported p1 gene sequences. An AGT trinucleotide variable-number tandem repeat was identified that ranged in copy number from 5 to 14 among the isolates. In addition, a novel p1 gene variant named 2c was identified in 6 of the isolates.
Journal of clinical microbiology 06/2011; 49(8):3000-3. · 4.16 Impact Factor
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ABSTRACT: The aim was to study the changes in the antimicrobial resistance of methicillin-resistant Staphylococcus aureus (MRSA) clones over a 5-year period (from 2000 to 2005) at a representative hospital in Beijing, China.
A total of 100 randomly selected MRSA strains were analyzed using antimicrobial susceptibility testing, pulsed-field gel electrophoresis, spa typing, multilocus sequence typing, SCCmec typing, and PCR for the Panton-Valentine leukocidin virulence factor.
Resistance to rifampin greatly increased from 32% (16/50) to 68% (34/50). High-level mupirocin-resistant isolates were found only in 2005, when four were identified. Intermediate susceptibly to quinupristin-dalfopristin increased from 22% (11/50) to 52% (26/50) between 2000 and 2005. The main antimicrobial resistance profiles changed from TC-GM-CI-EM-CM in 2000 to TC-GM-CI-EM-CM-RI in 2005. The main pulsed-field gel electrophoresis type changed from types C, L, and E in 2000 to types J, F, and N, respectively, in 2005. ST239-MRSA-III was the most predominant clone in 2000 and 2005, whereas ST5-MRSA-II was found only in 2005.
There were increasing levels of antimicrobial resistance and epidemiological changes in the hospital-associated MRSA strains isolated in this facility between 2000 and 2005.
Microbial drug resistance (Larchmont, N.Y.) 01/2011; 17(2):235-9. · 1.99 Impact Factor
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ABSTRACT: To obtain the genotype and antimicrobial susceptibility profiles of Campylobacter jejuni isolates from north China, 93 C. jejuni isolates (56 isolates from patients with diarrhoea, 7 isolates from Guillain-Barré syndrome patients and 30 isolates from chicken stools) were selected for multilocus sequence typing (MLST), PFGE and drug resistance testing. A total of 49 sequence types (STs) were identified from the entire panel of 93 C. jejuni isolates. Fifty-six isolates belonged to 14 clonal complexes, while 37 isolates could not be assigned to any known clonal complex. The most frequently observed clonal complexes were ST-21 (11 isolates), ST-353 (10 isolates) and ST-443 (6 isolates). Fifty-three PFGE SmaI patterns were identified among 93 isolates. No erythromycin-, gentamicin- or streptomycin-resistant isolates were found among the 44 strains isolated in 2008. Resistance to nalidixic acid, levofloxacin and ciprofloxacin was observed in 100 % (44/44) of the tested isolates. This study has shown the genetic characteristics of C. jejuni isolates in north China. In addition, overlapping clonal groups were defined by both MLST and PFGE for C. jejuni human and chicken isolates.
Journal of Medical Microbiology 10/2010; 59(Pt 10):1171-7. · 2.50 Impact Factor
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Maojun Zhang,
Qun Li, Lihua He,
Fanliang Meng,
Yixin Gu,
Minghuan Zheng,
Yunwei Gong,
Ping Wang,
Feng Ruan,
Lei Zhou,
Jing Wu,
Li Chen,
Collette Fitzgerald,
Jianzhong Zhang
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ABSTRACT: From June to July 2007, 36 cases of Guillain-Barre syndrome (GBS) occurred in a township in north China. Serological study and bacteria culture were performed to investigate the association between preceding Campylobacter jejuni infection and this GBS outbreak. Anti-C. jejuni antibodies were found in significantly higher numbers of GBS patients (IgM 84%, IgG 87.5%) than in healthy inspection cases (IgM 33%, IgG 27%). IgG anti-GM1 was the dominant anti-ganglioside antibody among the GBS patients. Seven C. jejuni isolates (four from human stool and three from poultry specimens taken from the patients' houses) were obtained. Serotyping and molecular analysis were used to investigate the genetic relatedness among these C. jejuni isolates. The four human isolates, collected from residents of the same district, were indistinguishable by both pulsed-field gel electrophoresis and multilocus sequence typing, suggesting these patients had a common source of infection. A new sequence type, sequence type-2993, was assigned to the human C. jejuni isolates, three of which belonged to Penner serotype heat-stable (HS):41. Both serotype and molecular subtype of the human C. jejuni isolates were different from those of isolates obtained from poultry specimens. Our results suggest that the antecedent C. jejuni infection triggered this GBS outbreak in China.
Foodborne Pathogens and Disease 05/2010; 7(8):913-9. · 2.26 Impact Factor
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ABSTRACT: Campylobacter jejuni ICDCCJ07001 (HS:41, ST2993) was isolated from a Guillain-Barré syndrome (GBS) patient during a 36-case GBS outbreak triggered by C. jejuni infections in north China in 2007. Sequence analysis revealed that the ICDCCJ07001 genome consisted of 1,664,840 base pairs (bp) and one tetracycline resistance plasmid of 44,084 bp. The GC content was 59.29% and 1,579 and 37 CDSs were identified on the chromosome and plasmid, respectively. The ICDCCJ07001 genome was compared to C. jejuni subsp. jejuni strains 81-176, 81116, NCTC11168, RM1221 and C. jejuni subsp. doylei 269.97. The length and organization of ICDCCJ07001 was similar to that of NCTC11168, 81-176 and 81-116 except that CMLP1 had a reverse orientation in strain ICDCCJ07001. Comparative genomic analyses were also carried out between GBS-associated C. jejuni strains. Thirteen common genes were present in four GBS-associated strains and 9 genes mapped to the LOS cluster and the ICDCCJ07001_pTet (44 kb) plasmid was mosaic in structure. Thirty-seven predicted CDS in ICDCCJ07001_pTet were homologous to genes present in three virulence-associated plasmids in Campylobacter: 81-176_pTet, pCC31 and 81-176_pVir. Comparative analysis of virulence loci and virulence-associated genes indicated that the LOS biosynthesis loci of ICDCCJ07001 belonged to type A, previously reported to be associated with cases of GBS. The polysaccharide capsular biosynthesis (CPS) loci and the flagella modification (FM) loci of ICDCCJ07001 were similar to corresponding sequences of strain 260.94 of similar serotype as strain ICDCCJ07001. Other virulence-associated genes including cadF, peb1, jlpA, cdt and ciaB were conserved between the C. jejuni strains examined.
PLoS ONE 01/2010; 5(11):e15060. · 4.09 Impact Factor
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ABSTRACT: Previous studies have implicated CagA [encoded by cytotoxin-associated gene A (cagA)] in Helicobacter pylori-associated gastroduodenal pathology and distinct subgenotypes of cagA may circulate in different pathological manifestations of cagA-positive H. pylori infection. To investigate cagA genotype and variants in Chinese H. pylori strains and explore their relationship with gastroduodenal diseases, the cagA status of 82 Chinese H. pylori strains was examined and variation in size of the 3' region of cagA in 71 of these strains was analysed by PCR. cagA was detected in 28 (100%) of 28 strains from peptic ulcer patients, two (100%) of two strains from gastric cancer patients, 32 (94.1%) of 34 strains from chronic gastritis patients and 17 (94.4%) of 18 strains from healthy volunteers. PCR products of the cagA 3' variable region were obtained from 71 (92.2%) of 77 Chinese H. pylori strains and could be classified into subgenotypes I, II and III, which gave PCR products of around 825, 900 and 950 bp, respectively. Subgenotype I cagA predominated in Chinese H. pylori strains (67/71), whereas subgenotype II cagA presented in two isolates from patients with chronic gastritis and subgenotype III presented in two isolates from healthy volunteers. Therefore, neither cagA nor its 3' region variants can be used as a sole marker for the presence of particular H. pylori-related gastroduodenal diseases in the Chinese population.
Journal of Medical Microbiology 04/2004; 53(Pt 3):231-5. · 2.50 Impact Factor
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ABSTRACT: To study the distribution of IS605, IS606 among clinical isolates of Helicobacter pylori in China.
A total of 104 H.pylori strains isolated from 5 different geographic regions in China were analyzed by PCR and dot-blot.
Forty-two strains out of the 104 isolates from 5 regions in China were found containing IS605 with 19 containing IS606. The frequency (66%) of IS605 positive strains from Yunnan province was higher than that from other areas. The different distribution of IS606 was neither associated with geographical regions nor with the presence of IS605 but IS606 were associated with the different clinical outcomes. However, the two reading frames ORFA and ORFB of IS605 were constantly coexisting.
In China, IS605 and IS606 of H. pylori were widely existing but the presence of IS605 in H. pylori might be associated with geographic origin.
Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 11/2002; 23(5):366-9.
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ABSTRACT: The cystic fibrosis transmembrane conductance regulator (CFTR) epithelial anion channel is a large multidomain membrane protein that matures inefficiently during biosynthesis. Its assembly is further perturbed by the deletion of F508 from the first nucleotide-binding domain (NBD1) responsible for most cystic fibrosis. The mutant polypeptide is recognized by cellular quality control systems and is proteolyzed. CFTR NBD1 contains a 32-residue segment termed the regulatory insertion (RI) not present in other ATP-binding cassette transporters. We report here that RI deletion enabled F508 CFTR to mature and traffic to the cell surface where it mediated regulated anion efflux and exhibited robust single chloride channel activity. Long-term pulse-chase experiments showed that the mature ΔRI/ΔF508 had a T1/2 of ∼ 14 h in cells, similar to the wild type. RI deletion restored ATP occlusion by NBD1 of ΔF508 CFTR and had a strong thermostabilizing influence on the channel with gating up to at least 40 °C. None of these effects of RI removal were achieved by deletion of only portions of RI. Discrete molecular dynamics simulations of NBD1 indicated that RI might indirectly influence the interaction of NBD1 with the rest of the protein by attenuating the coupling of the F508-containing loop with the F1-like ATP-binding core subdomain so that RI removal overcame the perturbations caused by F508 deletion. Restriction of RI to a particular conformational state may ameliorate the impact of the disease-causing mutation.
Journal of Molecular Biology.
