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To determine the distribution of two important virulence factors [lipooligosaccharide (LOS) and capsular polysaccharide (CPS)] inWhole-genome sequencing was carried out for 494Nine novel and 29 previously confirmed LOS classes were identified. LOS classes A, B, and C were the most common (48.2%, 238/494) among the 494 strains. Twenty-six capsular types were identified in 448 strains. HS2, HS4c, HS5/31, HS19, and HS8/17 were the most frequent CPS genotypes (58.7%, 263/448). Strains of 17 CPS genotypes (strain number5) had one or two prevalent LOS classes (Our results describe the genetic characteristics of the important virulence factors in

Few studies in China focused on serotypes of Streptococcus pneumoniae in patients with invasive pneumococcal disease (IPD). We aimed at investigating the serotype distribution for IPD-causing S. pneumoniae and vaccine coverage among Chinese children and adults. This was a multicenter, observational study to collect S. pneumoniae isolates from normal sterile sites and IPD-related clinical information among children and adults. Serotyping was performed by a Capsule-Quellung reaction test using type-specific antisera. The study collected a total of 300 eligible isolates (pediatric = 148, adult = 152) were serotyped in a central laboratory. The most prevalent serotypes were 19A (20.9%) and 23 F (20.3%) in the pediatric group; 3 (21.7%) and 19 F (11.8%) in the adult group. PCV10 had low-to-moderate serotype coverage rates for children (60.8%) and adults (34.2%). PCV13 and PPV23 had high coverage rates for children (89.9%, 93.2%) and adults (70.4%, 82.9%), respectively, Investigational PCVs including PCV15 and PCV20 had high estimated coverage rates in children (89.9%, 93.9%). The study identified 269 subjects with IPD reported as the primary diagnosis in the medical records. Sepsis (48/136, 35.3%) and pneumonia (48/133, 36.1%) had the highest occurrence in the pediatric and adult groups, respectively. Study findings showed that non-PCV7 S. pneumoniae 19A and 3 were the most prevalent serotypes in Chinese children and adults, respectively. High-valent vaccines had similar coverage rates and may have a greater potential in preventing IPD.

Objectives: The aims of this study were to analyse the serotypes of epidemic Haemophilus influenzae and changes in mechanisms of β-lactam resistance over the past decade. Results: Haemophilus influenzae isolates in Western Sichuan from 2013–2014 were non-typeable H. influenzae (NTHi). β-Lactam MICs for NTHi isolated during 2013–2014 were significantly higher than those from 2003–2004 (P < 0.05). Of 274 NTHi, 141 (51.5%) were β-lactamase-positive (TEM-1 type). There were 35 amino acid (AA) substitutions in ftsI among NTHi isolated from 2013–2014. However, NTHi isolates from 2003–2004 had only nine AA substitutions. Ordered multiple classification logistic regression analysis showed that different AA substitution patterns in ftsI had different effects on β-lactam MICs. The main factors affecting the ampicillin MIC were the mutations R517H (OR = 6.999), L389F (OR = 7.128), N526K (OR = 4.660) and D350N (OR = 0.450). The main factor influencing the amoxicillin/clavulanic acid MIC was an N526K mutation (OR = 9.349). The main factors affecting the cefuroxime MIC were the mutations S357N (OR = 37.453) and N526K (OR = 14.816). Compared with 2003–2004, gBLNAR and gBLPAR isolated from 2013–2014 increased significantly from 13.0% (7/54) and 9.3% (5/54) to 38.2% (84/220) and 45.5% (100/220), respectively (P < 0.001). In the ‘others’ group of ftsI gene mutations, 13 NTHi had the same ftsI gene mutation pattern and 24 AA substitutions. Conclusion: These results confirm that β-lactam-resistant NTHi isolates increased rapidly. AA substitutions in ftsI were more complex and diversified in 2013–2014.

To compare the pathogenicity of isolates of sequence type 7 (ST-7)Four ST-7Among the four ST-7The differences in the capsular structure of the four isolates of ST-7

The carriage rate and serotype distribution of Streptococcus pneumoniae (S. pneumoniae) in a healthy population in China remains unclear. In this study, we collected the oropharyngeal swabs from 513 individuals in Xinjiang, China. Real-time PCR targeting the lytA gene and 12 serotypes were assessed to identify S. pneumoniae carriage. The total carriage rate of S. pneumoniae was 70.4% (361/513). The most prevalent serotypes were 19B/F, 18B/C, 5, and 6A/B. The highest carriage rate of S. pneumoniae was noted in children aged 6-10 years (88.6%), which merits further attention. The co-colonization rate of two or more S. pneumoniae serotypes was 79.8% (264/331). This study aimed to investigate the baseline pneumococcal carriage rate among healthy individuals in China to improve our understanding of the epidemiology of S. pneumoniae.

To compare the detection effect of Legionella pollution in spring water by three methods, namely traditional plating method, fluorescent quantitation PCR method and ethidium monoazide (EMA) fluorescent quantitation PCR method.Every month (except May), we collected 11 water samples from the 5 selected hot spring pools in one hot spring resort in Beijing in 2011. A total of 121 water samples were collected, and then were detected by the above three methods qualitatively and quantitatively.In our study, the Legionella pollution rate was separately 74.4% (90/121), 100.0% (121/121) and 100.0% (121/121) by the above three methods. The quantitative value of Legionella in the 121 water samples detected by the three methods were around 0.10-216.00 colony-forming units (CFU)/ml, 1.47-1557.75 gene units (GU)/ml and 0.20-301.69 GU/ml, respectively. The median (25th and 75th percentiles) was 75.30 (32.51-192.10) GU/ml, 36.46 (16.08-91.21) GU/ml and 5.30 (0.00-33.70) CFU/ml, respectively. The difference in the quantitative value of Legionella detected by the three methods showed statistical significance (χ(2) = 187.900, P0.01). The quantitative value of Legionella detected by fluorescent quantitation PCR method was the highest, followed by the value Legionella detected by EMA-fluorescent quantitation PCR method and traditional plating method.The sensitivity of the PCR methods was higher than traditional plating method, in detecting Legionella pollution in spring water, especially the EMA- fluorescent quantitation PCR method, which was more suitable for detecting Legionella in water.

To analyze the levels of human serum antibody against Neisseria meningitidis serogroup C measured by serum bactericidal assay (SBA) and ELISA.SBA and a modified ELISA were applied to measure the serum bactericidal titer and the specific concentration of immunoglobulin G (IgG) against meningococcal serogroup C in sera samples. Seventy-five sera were from healthy adults without undertaking vaccination while another 429 and 388 pre- and post-vaccinated sera were from 143 infants and 194 young children immunized with conjugate vaccine or polysaccharide vaccine, respectively. Correlation between serum bactericidal titer and the concentration of specific IgG against meningococcal serogroup C was analyzed.The concentration of meningococcal serogroup C specific IgG in healthy adults showed a strong correlation (r=0.814 33, P0.001) with serum bactericidal titer through linear regression analysis. Weak correlation was observed between SBA titers and IgG concentration in pre vaccinated sera of infants and children (conjugate/polysaccharide vaccine) (infants: r=0.140 64, P0.100/r=0.2899, P0.05; children: r=0.540 40, P0.05/r=0.194 36, P0.05). After immunization with 2-dose conjugate vaccine in infants and 1-dose in children, a strong correlation between the two panels of results was observed (r=0.809 38, P0.001 and r=0.837 23, P0.001 respectively). However after immunization with polysaccharide vaccine, the correlation between serum bactericidal titer and concentration of specific IgG was weak (r0.50000).Among healthy adults and post vaccinated infants or young children immunized with conjugate vaccine, the concentration of specific IgG was comparable to the serum bactericidal titer against meningococcal serogroup C. However, it was not unfavorable to use ELISA as the principal means of measuring serum antibody responses to polysaccharide vaccine for infants under 1 year old.

To investigate the contamination state of Legionella in cooling water samples from different places in Wuxi city and reveal the molecular biological characteristics of Legionella strains.112 parallel water samples (500 ml each) were collected from 56 sites in Wuxi city during year 2009 - 2010. The samples were used for Legionella test and quantitative culture. The isolated Legionella strains were used for serotyping, pulsed-field gel electrophoresis (PFGE), sequence-based typing (SBT), and intracellular growth were tested.The positive proportion of Legionella was 39. 3% (22/56) among all sampling sites. A total of 29 Legionella strains were isolated, and the serotypes include LP1, LP3, LP5 and LP6. LP1 serotype was the major one with a proportion of 65.5% (19/29). 29 Legionella strains got 17 PFGE types. There were 10 SBT types among 10 Legionella strains with different PFGE types. Comparing to LP1 strain (ATCC 33152), WX2011062 (LP6) and WX2011067 (LP5) had strong intracellular growth ability in mouse peritoneal macrophages J774 cell line (the amount of intracellular bacteria on day 0 after infection were (5.5 +/- 1.32) x 10(5), (3.9 +/- 0.60) x 10(5), (7.8 +/- 0.76) x 10(5) CFU/ml, respectively; the amount of intracellular bacteria on day 3 after infection were (58.3 +/- 1.61) x 10(5), (2700.0 +/- 655.74) x 10(5), (3066.7 +/- 208.17) x 10(5) CFU/ml, respectively).The Legionella contamination existed in cooling water samples from different places in Wuxi city. Legionella strains isolated showed high genetic variation. Some Legionella strains had vigorous intracellular growth ability.

To characterize the meningococcal strains isolated from cases and close contacts with meningococcal disease associated with an outbreak in a jail in May 2010 by investigating the national distribution of hyperinvasive ST-4821 serogroup C clone associated with this outbreak.The cases were described based on the clinical symptoms and laboratory results. Pharyngeal swabs were cultured for N. meningitidis from men in the jail. Meningococcal isolates were identified by serogrouping, pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST), respectively. Four hundred and sixteen serogroup C N. meningitidis strains were collected from 27 provinces between 2003 and 2010 for a nationwide survey and analyzed by PFGE and MLST.Three persons in a jail system were infected with invasive N. meningitidis serogroup C. All isolates tested had matching PFGE patterns and belonged to the multilocus sequence type (ST) 4821 clonal complex. All 47 N. meningitidis strains were identified from the pharyngeal swabs of 166 peoples in the jail, and 26 of them belonged to ST-4821 serogroup C clone, and 90.14% (375/416) serogroup C strains identified in the nationwide survey belonged to the ST-4821 complex. The ST-4821 serogroup C clone was spread nationwide, distributed in 24 provinces, especially in eastern provinces between 2003 and 2010.Endemic transmission and carriage rate of ST-4821 serogroup C clone are high in this jail system. The ST-4821 serogroup C clone is spreading in China and nationwide distributed despite the existence of some effective vaccines.

To analyze the characteristics of Sequence-based Typing (SBT) of the Serotype 1 Legionella pneumophila (Lp1) isolated from environmental water in China, and then create a preliminary database.A total of 82 strains of Lp1 isolated from environmental water in 9 provinces of China between 2005 and 2008 were genotyped by SBT method and Pulsed-field Gel Electrophoresis (PFGE) method. The results of the two different typing methods were then compared by cluster analysis, adopting BioNumerics version 5.1 software.By SBT method, the 82 strains of Lp1 were divided into 22 ST types, of which 17 new types and one new allele was discovered. The dominant type was ST-1 type, found in 8 provinces, accounting for 46.3% (38/82). ST-1, ST-150, ST-154, ST-159, ST-160 and ST-630 types were found in more than 2 isolated-sites; while more than 2 different ST types were found in 5 isolated-sites, as site B4, B5, B6, S3 and S8. In cluster analysis, 15 ST types were grouped into three complexes (ST-1 complex, ST-154 complex and ST-149 complex); and the other 7 ST types were not assigned complex. By PFGE method, 46 banding patterns were observed. As a result of the combination of the two methods, the 82 isolates strains could be divided into 54 molecular types, which showed a reliable accordance in the cluster analysis between the two methods.The SBT of the Lp1 in environmental water in China was unique. From the study, a preliminary SBT database was set up.

To analyze the molecular types of Legionella (L.) pneumophila strains isolated in China, and to develop the PulseNet-China Database of L. pneumophila.Pulsed field gel electrophoresis (PFGE) was used to analyze 262 L. pneumophila strains collected from 11 provinces between 2004 and 2009 in China. Different kinds of genomic DNA in different L. pneumophila strains were isolated and separated after digesting with Asc I. BioNumerics software was used to analysis the PFGE fingerprints.L. pneumophila strains isolated in China were quite different regarding their PFGE patterns. There were 108 PFGE types among the 262 strains tested in this study. The similarity value of these strains was in the range of 16% - 100% and the same types were discovered in different provinces and years.L. pneumophila strains isolated in China were with high genetic variations. There might be different clones existed in China. The development of PulseNet China Database was thus of great significance in monitoring the L. pneumophila strains in the future.

To type Klebsiella pneumonia through methods including pulse-field gel electrophoresis (PFGE) in combination with multilocus sequence typing.Four selected different EPs, referring to the Standard Operating Procedure of PulseNet China, were used. The single colony of Klebsiella pneumonia was quantified after enriched culture. Embedding organisms in agarose and genome DNA were lysed with Proteinase K and then digested by restriction endonuclease XbaI, to produce agarose gel. Fingerprint was obtained through PFGE and bands were marked with their molecular weights and then analyzed by BioNumerics software. Using MLST to analyze the strains that were highly similar, by PFGE typingBy comparing the four results from each EPs, fk3 (switch time from 6s to 36s, total run time is 18.5 hours) seemed to be better than the others. 59 strains of Klebsiella pneumonia were divided into 47 PFGE types and 19 PFGE clusters. The highly similar strains could be typed into ST-340, ST-342, ST-343, ST-344, ST-345 by MLST. Among them, ST-342, ST-343, ST-344, ST-345 types were all new MLST types that were reported in China.Highly similar Klebsiella pneumonias typed by PFGE could also be typed by MLST.

To investigate the genotypic characteristics and persistence of Legionella pulsed-field gel electrophoresis (PFGE) patterns in 16 air-conditioner cooling towers in six different public sites of Shanghai.From May to October, continuous sampling was operated once per month in 2007. Legionella strains isolated from the 16 cooling towers were confirmed by serological and latex agglutination. PFGE was applied for the fingerprinting of the isolates, while the cluster results of PFGE were analyzed by BioNumerics software.131 strains of Legionella were isolated, including L. pneumophila, L. bozemanae, L. micdadei and L. anisa. 52 distinguishable PFGE patterns were differentiated among the 16 cooling towers, with 37 patterns were owned by just one cooling tower, which was not shared with other cooling towers, while 15 patterns were shared by more than 2 cooling towers. All the cooling towers had ≥ 2 PFGE patterns, while in 13 cooling towers the same PFGE patterns were recovered during the six months. From June to October of 2007, 18 strains of Legionella belonging to the PFGE pattern of LPAs.SH0078 were isolated continuously from 6 cooling towers.This study demonstrated great genotypic diversity and complexity of Legionella in cooling towers. Persistence of the PFGE patterns was observed in 81.25% of the cooling towers. The PFGE pattern of LPAs. SH0078 was distributed widely, suggesting it might be the dominate strain in Shanghai.

Meningococcal disease, caused by Neisseria meningitidis (Nm), is still one serious threatening infectious disease with high mortality. Vaccination is available for prevention and control of such disease. Based on the chemical structure of capsule polysaccharide, Nm strains were classified into 13 serogroups. Meningococcal polysaccharide vaccines and polysaccharide conjugated protein vaccine against serogroup A, C, W135 and Y were efficacious and have been widely used. Because of poor immunogenicity and the structurally homologous with neural cell, capsule polysaccharide of serogroup B Nm can not be used as vaccine candidate. In last several decades, B group vaccines develoment focused on the proteins research. Based on the out membrane protein and reverse vaccinology technology, progress of B group vaccine were accelerated. Several meningococcal B vaccine showed favorable immunogenicity and efficacity. Some B vaccines have been licensed and widely used.

To establish a method based on PCR for serotyping of Streptococcus pneumonia isolates. PCR serotyping method was applied for investigating the serotypes of S. pneumonia strains.12 pairs of primers targeting different serotypes of S. pneumonia were designed and synthesized. After optimizing the PCR amplification reaction, sensitivity and specificity of each pair was performed. We applied the PCR methods for testing the serotypes of the isolated S. pneumonia strains.Each pair of primers showed satisfied PCR sensitivity and specificity. Of all 119 S. pneumonia strains tested by PCR serotyping method, 113 isolates were identified (3, 5, 6A/B, 9A/V, 14, 18, 19A, 19F, 23F) with 6 isolates were unable to be serotyped.We developed a simple, reliable and economic method for S. pneumonia serotyping which could be used for testing the serotypes of S. pneumonia that had been prevailed among general population.

Haemophilus influenzae (Hi) is a pathogen exclusively found in humans. It causes a wide range of infections from the upper respiratory tract to serious invasive diseases. Such as pneumonia, septicemia and meningitis. Strains of Hi are usually classified into six serotypes a to f and nontypeable H. influenzae (NTHI) according to the antigenicities and compositions of their polysaccharide capsules. Hib was a common cause of serious infections in younger children. The polysaccharide-protein conjugate vaccines against Hib had almost eliminated H. influenzae as a cause of pediatric meningitis. However, NTHI remains an important pathogen, particularly in children and the elderly. Efforts to understand and control NTHI disease have been hampered by the diversity of these bacteria. This review introduced the study progress about pathogenic mechanism of NTHI. In order to provide the help for development of vaccine, clinic treatment and prevent the occurrence of diseases causing by NTHI.

To research the relationship between variable number of tandem repeats sequences (VNTRs) in genome and serogrouping of Neisseria meningitides (N. meningitidis).118 meningococcal strains which were isolated from invasive patients and carriers in China were serogrouped by antiseria agglutination testing. 38 strains were serogroup A. 80 were serogroup C. And then four variable number of tandem repeats sequences (VNTRs) loci in the DNA of 118 meningococcal strains were amplified by PCR. A dendrogram was constructed on the basis of DNA length of PCR productions by the Unweighted Pair Group Method with Arithmetic Averages (UPGMA). The two pair primers-specific were designed based on the nucleotide variant afterward the VNTR01 locus (T01-A and T01-AC, T01-C and T01-AC) to amplify 118 meningococcal strains.118 strains were classified into five VNTR groups. 98.3% (116/118) strains were identical between serogrouping and VNTRs genotyping. 97.3% (36/37) strains of serogroup A were categorized into group I of VNTRs genotyping. 98.7% (75/76) strains of serogroup C were categorized into group II of VNTRs genotyping. According to the presence or absence of PCR products using two pair primers-specific, 118 strains were classified into two groups which correspond to serogroup A and C. The sensitivity of primers-specific T01-A and T01-AC was 89.5% (34/38). The specificity of primers-specific T01-A and T01-AC was 100% (38/38). The sensitivity of primers-specific T01-C and T01-AC was 100% (80/80). The specificity of primers-specific T01-C and T01-AC was 95.2% (80/84).VNTRs genotyping has high relation to serogrouping A and C. The nucleotide sequence variations in the galactosyl-transferase gene related to the serogrouping A and C.

To optimize the serum bactericidal assay (SBA), detect and analyze the bactericidal antibody level against Neisseria meningitidis serogroup C strains after divalent polysaccharide (A plus C) vaccine immunization.Two Neisseria meningitidis serogroup C strains, vaccine candidate strain (C11) and epidemic strain (053442), were selected as targets. The national Neisseria meningitidis standardized serum was used as reference serum. Pel-Freez infant rabbit complements was available. The optimized SBA method was used to detect bactericidal antibody against strain C11 and 053442 for 122 pairs of sera before and after immunization with a divalent polysaccharide (A and C) vaccine.The strain C11 and 053442 both could be used as targeted strain for SBA. The optimized concentration of targeted strain was achieved when a whole-cell suspension of 0.35 A at 600 nm was diluted 4 x 10(4) times. Before immunization, SBA geometric mean titers (GMT) of 122 sera against strain C11 and 053442 were 1:1.75 and 1:2.63 respectively, and the protective rates were 9.8% and 17.2% respectively. After immunization, the GMTs and the protective rates of 122 sera both rose significantly (P0.01), the GMTs against strain C11 and 053442 were 1:483.73 and 1:412.57 respectively. The protective rates against strain C11 and 053442 were 100% and 95.9% respectively.Immunization with a divalent polysaccharide (A and C) vaccine could elevate remarkably the population SBA titer against Neisseria meningitidis serogroup C strains of different subtypes, but the surveillance of vaccine effect against different targeted strains remains necessary.

To research the distribution and molecular epidemiology of insertion sequence IS1301 in Neisseria (N.) meningitidis strains in China, so as to provide scientific and available evidence for a new method of genotyping in N. meningitidis strains with IS1301.Examined the IS1301 by PCR in 219 N.meningitidis strains from 16 provinces and 3 cities during 2007 and 2008 in China, productions of amplification were sent for sequencing. The positive N. meningitidis strains were analyzed by pulse field gel electrophoresis (PFGE) and nucleic acid blotting hybridization(Southern blot) by electrophoresis.The positive rates with IS1301 were 15.53%, 11.11%, 20.75%,6.17% and 28.57% for four serotypes (A, B, C, N) respectively. The sequence comparability between the amplification productions and No.Z49092.1 N. meningitidis which registered in GenBank was 94%-100%. There were two types of clusters divided by cladogram analysis. There appeared large IS1301 sequence difference between the serotype C and others. The number of IS1301 replica ranged from 6-17 per strain at least. The number of IS1301 replica changed in the same type of PFGE N. meningitidis respectively.Typing by IS1301 combined with PFGE could comprehend the homology and genetic polymorphism of N.meningitidis epidemic strains at the molecular level.

Two hundred and seventy-three Haemophilus influenzae strains isolated from pediatric pneumonia patients in China were studied. We used Multilocus Sequence Typing (MLST) to analyze genotypic characteristics. All strains were biotyped and serotyped. Relatedness and patterns of genes among isolates were determined by the analysis of MLST and eBURST. H. influenzae primarily causes acute pneumonia in children under 1 year old. Nontypeable H. influenzae was responsible for most cases of pediatric pneumonia. All 273 strains were classified into eight biotypes. They mostly belonged to the I, II, and III biotypes (17.6%, 43.6%, and 22.7%, respectively). 62 strains (22.7%) produced beta-lactamase. We found 28 novel alleles. Fifty different STs were found by MLST, of which 39 were novel. These were ST477 through ST508 and ST521 through ST527. Group 17 and predicted founders 503 were new groups in this study. No STs correlated with strains from Korea, which is adjacent to China. The H. influenzae strains from China appeared to have heterogeneous ST types patterns which may be the reason no outbreaks or epidemics of H. influenzae infections have occurred in Chengdu city, Sichuan, China.

To develop a rapid method for detecting Haemophilus influenzae by multiplex polymerase chain reaction (M-PCR).Primers (Hi) were designed for amplification of p6 gene coding P6 protein of Haemophilus influenzae, which was used to identify Haemophilus influenzae species. Primers (Hi-cap) were designed for amplification of bexA gene which coding capsular polysaccharide (cap) synthesis was used for detecting whether Haemophilus influenzae isolates possess bexA gene relating to cap synthesis. Twelve primers (Hia-Hif) were designed for amplification of cap synthesis gene to identify the cap-type of Haemophilus influenzae. Other relative enteric pathogenic bacteria were amplified by M-PCR to serve as controls. 200 strains isolated from patients were identified. Results from M-PCR were compared to two methods including V and X factors grow requirement test and standard slide agglutination serotyping (SAST).The results indicated that the M-PCR assay was high specificity and sensitivity and might be valuable for differential diagnosis of Haemophilus influenzae. The sensitivity of detection was 0.935 pg. 189 strains out of the 200 belonged to Haemophilus influenzae isolates, and one isolate was cap-type f. An agreement results were seen among the V and X factors grow requirement test, SAST and M-PCR methods.M-PCR method showed satisfactory sensitivity, specificity and stability for detecting and identifying Haemophilus influenzae, and could be used in clinic diagnosis, surveillance and rapid diagnosis for plague of Haemophilus influenzae.

To study the characteristics of epidemiology and molecular typing on Neisseria meningitidis serogroup C strains associated with outbreaks of Anhui province and sporadic cases in China, using pulsed field gel electrophoresis (PFGE).212 Neisseria meningitidis serogroup C strains were isolated from invasive meningococcal cases, close contacts and healthy carriers, including 48 strains from Anhui province with 38 strains associated with serogroup C outbreaks. PFGE were performed by genomic DNA digestion with Nhe I restriction enzyme. The results of PFGE were analyzed by BioNumerics software (Version 4.0, Applied Maths BVBA, Belgium).A total number of 212 Neisseria meningitidis serogroup C isolates were typed by 43 patterns, named AH1 to AH43. In China, AH1 pattern was the major PFGE pattern with 69.3% (n = 147) of all strains, distributed in 11 provinces. Three types of PFGE patterns (AH1 to AH3) were found in 48 strains from Anhui province, in which, 93.8% (n = 45) belonged to AH1. 97.4% (n = 37) of 38 strains associated with serogroup C outbreaks in Anhui province showed AH1 pattern. A total of 53 serogroup C strains were isolated from invasive meningococcal cases with 67.9% (36/53) of AH pattern. 71.9% (87/121) of serogroup C strains isolated from contacts of invasive meningococcal cases was AH1 pattern and 63.2% (24/38) of the strains from healthy carriers showed AH1 pattern.By PFGE typing and analysis, AH1 pattern of Neisseria meningitidis serogroup C strains was proved to be the main clone which causing the outbreaks in Anhui province and might be responsible for the sporadic serogroup C meningococcal disease epidemics else where in the country.

The aim of this study was to characterize the molecular features of serogroup C Neisseria meningitidis strains circulating in Beijing, China. Twenty out of 23 strains belonged to ST 4821. The causative serosubtype for meningococcal meningitis was P1.12-1,16-8. All of the strains expressed class 3 PorB protein. Among the five pulsed-field gel electrophoresis patterns observed, pattern III predominated.

We developed a multiplex polymerase chain reaction (PCR) method for detection, identification and sero-grouping strains of N. meningitidis.The gene of crgA was selected for detection and identification of N. meningitidis and the 230 bp of crgA fragments were amplified. The genes of siaD and orf-2 were used for sero-grouping. With this technology, N. meningitidis of serogroup A,B,C,Y and W135 can be differentiated from the specific fragment 400 bp,450 bp,250 bp, 120 bp,120 bp.This multiple PCR method seemed to be more sensitive than latex agglutination. We used this method to detect 61 isolates of N. meningitidis and the 230 bp of crgA fragments were amplified. No positive results were obtained from on 4 strains of non-N. meningitidis. It seemed that the PCR method which was based on crag, was specific in detecting N. meningitidis. The 55 strains of N. meningitidis could be grouped into A,B,C,Y and W135 by this multiplex PCR. No gene fragment was synthesized for the other serogroups of N. meningitidis. No cross-reaction was observed for other bacterial species. 4 samples of meningococcal meningitis patients were detected by the multiplex PCR and two of them were positive with 400 bp band,representing serogroup A of N. meningitidis but all were negative to culture and latex agglutination. 18 samples of Japanese (B) encephalitis patients were also detected by multiplex PCR and no positive result was obtained which was consistent to the finding from culture and latex agglutination.This multiplex PCR method showed its potential in clinical diagnosis and epidemiological investigation.

A suspicious meningococcal meningitis death case was reported to the Beijing CDC. The blood specimen was analyzed via multi-PCR and MLST. 6 isolates from close contacts were analyzed via PFGE and MLST. According to the results of the above analyses, the cause of this case was identified as a serogroup A Neisseria meningitidis, which, in terms of sequence typing, belonged the ST7 group.