Molecular characterization and drug-resistance of Mycobacterium tuberculosis strains in Xuzhou, China.

To understand the genetic diversity and drug resistance status of Mycobacterium 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-loci 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.

[Analysis on pathogen detection for 53 influenza outbreaks].

OBJECTIVE: In order to provide a scientific basis for influenza prevention and control, analyzing the epidemic characteristics and laws of influenza outbreaks in Xuzhou area during 2005-2011. METHOD: Using fluorescent-PCR method to detect influenza virus nucleic acid on Nasopharyngeal swab specimens collected from influenza outbreak cases during 2005-2011 and fast classifying influenza virus A1 (H1N1), A3 (H3N2), new H1N1 BV (Victoria) and BY (Yamagate) on subtypes. At the same time, isolating the influenza virus with MDCK cells, and sending them to the National Influenza Center for review, after the preliminary identification of the isolated influenza virus. RESULTS: During 2005-2011, there are 53 influenza outbreaks in Xuzhou area, which caused by influenza virus subtype BV accounting for 26.42% (14/53), A3 accounting for 49.1% (26/53), A3 and A1 mixture accounting for 3.77% (2/53) and the new H1N1 accounting for 20.75% (11/53). The outbreaks in 2007 and 2009 mainly caused by A3, and show that the winter spring (January) and summer autumn (September) as two popular peaks during 2005-2011; BV mainly causes the outbreaks from Feb. to Jun. CONCLUSION: In Xuzhou area, since the winter of 2005, influenza virus subtype BV, the A3, and new H1N1 has alternately as mainly predominant strain, caused local influenza outbreaks. In which BV has increased trend year by year during 2005-2011. The students in primary and secondary schools are the major crowd of influenza outbreaks. Fluorescent-PCR detection methods could be a preferred method for reliable and rapid diagnostic of epidemic influenza outbreaks.

[Etiological analysis of influenza surveillance data in Xuzhou from 2005 to 2011].

OBJECTIVE: To investigate the prevalence and subtypes of influenza viruses in Xuzhou city from 2005 to 2011 and to provide the scientific supports for influenza prevention and control in this religion. METHODS: The throat swab samples were collected from the influenza-like cases from national influenza like illness sentinel hospital in Xuzhou. The samples were used for influenza virus isolation and identification, sent on the national flu center to confirm according to the "national influenza surveillance program" and "influenza virus and experimental technology". RESULTS: From Oct. 2005 to Dec. 2011, a total of 9561 swab specimens were collected in which 1152 strains were identified for influenza viruses with total isolated rate of 12.0%. Among these strains, 708 strains were A1 (H1N1) subtype (14.2%), 466 strains were A3 (H3N2) subtype (40.5%), 78 strains were new H1N1 subtype (6.8%), 362 strains were BV (Victoia) subtype (31.4%) and 82 strains were BY (Yamagate) subtype (7.1%). The top detection rate (25.9%) arose in 2007, secondary detection rate (17.4%) occurred at 2009 and the lowest one (2.3%) appeared in 2011. From the winter of 2005 to the spring of 2006 A1 (H1N1) subtype had appeared as predominant strains but in the winter of 2006 the predominant strains were BV subtype. It changed to A3 subtype in 2007 to 2009 and the other three dominant strains were A1, BV and BY in 2008. In the winter of 2009, both A3 (H3N2) and new H1N1 subtype were predominant strains. BV subtype was predominant strains in 2010 to 2011. The prevalence of A3 subtype appeared in all the year while prevalence of BV only arose in the spring and winter. So the detection rate was high in January (34.4%) but low in August (2.2%). The influenza population is correlated with age, the highest detection rate arose in 5-age group and the lowest detection rate appeared in 25-age group. CONCLUSION: Influenza subtype A1, A3, New H1N1 are all appeared as predominant strains in Xuzhou city from 2005 to 2010. Besides, the prevalence of BV subtype is stronger in recently.

[Research on the aetiology of suspected enterovirus infected patients in Xuzhou district in 2009].

OBJECTIVE: To understand and master the situation in which enterovirus caused hand-foot-and-mouth disease (HFMD) in Xuzhou district in 2009 so as to provide scientific basis for the control and prevention of hand-foot-and-mouth disease. METHODS: The researchers adopted fluorescence RT-PCR method to detect EV and EV71 as well as the CA16 specificity RNA from 222 samples of anal swabs and oropharyngeal swabs from the 240 cases who were diagnosed clinically as hand-foot-mouth disease infected by enterovirus. Also, the researchers conducted EV71-IgM antibody detection on 114 samples of acute phase serum with ELA method. RESULTS: Among the 240 enterovirus infected patients, the total EV infection rate is 72.50% (174/240), among which EV71 infection rate is 57.92% (139/240), CoxA16 infection rate is 9.17% (22/240), and other EV infection rate is 5.42% (13/240). The EV71-RNA positive rate of the samples of 222 anus swabs among the 240 suspected enterovirus infected patients is 45.94% (102/222), the samples of swallow swab EV71-RNA positive rate is 25.68% (57/222) and the EV71-IgM antibody positive rate of 114 samples of acute phase serum is 86.84% (99/114). The EV71-RNA positive rate of oropharyngeal swabs of 254 healthy children is 1.57% (4/254) , and no CoxA16-RNA was detected. In the oropharyngeal swabs of 54 close contacts (medical personnel), the EV-RNA detected is negative. The positive rate of EV71-IgM antibody of the 258 healthy children's serum samples is 2.71% (7/258). CONCLUSION: The widespreading of hand-foot-mouth disease in Xuzhou district is caused mainly by type 71 enterovirus. Inapparent infection of type 71 enterovirus exists among children under the age of 3 during the time of widespreading period and IgM antibody develops in them. It is difficult for adults to be infected by EV71 even if they contact the contagion source closely. The positive rate of EV71-IgM antibody in the samples of acute phase serum of suspected cases is the highest (86.84%), and the second highest is the positive rate of RNA of EV71 of anal swabs (45.94%) and of the EV71 of oropharyngeal swabs (25.68%). ELA reagent kit is used in the early diagnosis of EV71 infection for it is easy to operate, fast and economic, so, it is worth popularizing in the grass-root medical units.