1.Clinical application of PCR and high resolution melting analysis for rapid identification of Brucella isolates
Xueni LIANG ; Buyun CUI ; Lingling MAO ; Wei REN ; Jingbo YU ; Wencheng XUE ; Dongya MENG
Chinese Journal of Zoonoses 2015;(3):255-259
The aim of this study is to develop a rapid and accurately species typing method for Brucella isolates by using High Resolution Melting (HRM ) analysis .Six pairs of primers were used according to the reference for the sequence of pur‐pose gene .Nineteen biotypes of six species Brucella standard strains were identified by PCR‐HRM analysis and this analysis was used to detect the 35 clinical isolates .Results showed Brucella amplified specific melting curves were different from con‐trasted strains with primer Bspp .The six species Brucella standard strains have own characteristic curve shape from each oth‐ers by PCR‐HRM analysis with five pairs of primers .Thirty‐five clinical isolates of Brucella have entirely consistent with PCR‐HRM curve shape with Brucella melitensis standard strains .So ,PCR‐HRM analysis methods can accurately identify Brucella strains ,especially clinical isolated Brucella melitensis ,and may be used in clinical microbiology laboratories .
2.Bacillus anthracis isolates analysis by multiple-locus variable-numbers of tandem repeats analysis,Liaoning,China
Lingling MAO ; Jiang TIAN ; Lu LEI ; Xuesheng LIU ; Wei ZHANG ; Meimei ZHANG ; Yue HAN ; Wenqing YAO
Chinese Journal of Zoonoses 2015;(3):232-234
The epidemic characteristics and genotype of Bacillus anthracis strains in Liaoning Province ,China was analyze in this study .Six Bacillus anthracis strains from 2001 to 2011 were studied with multiple‐locus variable‐number tandem repeat analysis (MLVA) .BioNumerics4 .0 software was used to analyze the DNA fingerprint of statistics ,and cluster analysis results were obtained .Clustering analysis found that the 6 strains could be divided into two genotypes .For anthrax outbreaks ,the ge‐netic markers of multiple‐locus variable‐number tandem repeat were highly similar .It's suggested that MLVA is quite useful for investigation of strain relatedness in regions of outbreaks .
3.Prevalence of anti-HBV antibody among immunized population and evaluation of different detection methods of anti-HBc antibody
Xiaoli ZHANG ; Jinlong WANG ; Guanghua LIN ; Yingping CAO ; Jianlin ZHOU
Chinese Journal of Zoonoses 2015;(3):289-292
In this study ,we detected the positive rate of anti‐HBs and anti‐HBc antibody among the subject population in Fujian Medical University Union Hospital ,and to evaluate different detection methods of anti‐HBc antibody .The positive rate of anti‐HBs and anti‐HBc antibody were detected by chemiluminescent microparticle immunoassay (CMIA) and one‐step com‐petitive enzyme‐linked immunosorbent assay (ELISA) from the year 2012 to 2013 .The subject population was divided into three groups :group 1 with the age of less than 2 years old ,group 2 with the age of 2‐20 years old ,and group 3 with the age of more than 20 years old .The positive rates of anti‐HBV antibody in the different groups were analyzed .Furthermore ,anti‐HBc antibody of 92 samples selected from the immunized population was detected by CMIA and three kinds of ELISA reagents . Meanwhile ,the detection of anti‐HBc antibody by the same ELISA reagent but different operating modes were performed in these samples .The highest positive rate of anti‐HBs antibody was detected in group 1 ,and there was no significance difference of positive rate between two detection methods of anti‐HBs antibody among three groups .The positive rate of anti‐HBc anti‐body using CMIA was significantly lower than those with ELISA among group 1 and 2 .Among the 92 samples ,the positive rate of anti‐HBc antibody was 2 .2% using CMIA .With three kinds of method of ELISA reagent ,the positive rate of anti‐HBc antibody were 79 .3% ,82 .6% and 94 .6% ,respectively ,and there was no statistical significance among the results of three ELISA reagents .Anti‐HBc was not detected from 19 samples using ELISA methods with different operating modes .It's con‐cluded that the anti‐HBs antibody declined with the increase of age ,and it is necessary to discriminate the specific population to strengthen immune system .The obviously higher positive rate of anti‐HBc antibody was found by ELISA in immunized popula‐tion than that by CM IA . Concerning on the false positive of ELISA , specimen sampling with one specific test item or the CMIA method was recommended to detect the anti‐HBc antibody .
4.Investigation of Toxoplasma infection among planned pregnant women in Chongqing,China
Jun LIU ; Qing CHEN ; Liu YANG ; Yang HE ; Qi TONG
Chinese Journal of Zoonoses 2015;(3):284-288
We investigated the Toxop lasma infection prevalence among planned pregnant women in Chongqing ,and to pro‐vide reference for the first level intervention of birth defects in the region .A total of 11 953 planned pregnant women were se‐lected by proportionally stratified multi‐stage random sampling method .Questionnaire survey was given to the women ,and blood samples were collected .Specific IgM and IgG antibodies against Toxoplasma were detected with ELISA .Results showed that among the 11 953 cases surveyed ,Toxoplasma IgM was positive in 71 cases ,with the positive rate of 0 .59% ;IgG was positive in 771 cases ,with the positive rate of 6 .54% .The positive rate of IgM and IgG antibodies in the metropolitan core re‐gion of the city was higher than that in the suburb areas (χ2 =35 .28 ,P<0 .000 1 ;χ2 =82 .65 ,P<0 .000 1) .The positive rate of IgM antibody increased with the educational level (χ2trend=3 .25 ,P=0 .001 1) .The positive rates of IgM and IgG varied in occupations among women (χ2 =13 .93 , P= 0 .016 0;χ2 = 15 .58 ,χ2 =0 .008 1) ,with the highest rate of public officials . Planned pregnant women with the history of abnormal pregnancy outcomes had higher positive rate of T .gondii IgM and IgG antibodies than those in the control (χ2 =6 .85 ,P=0 .008 9;χ2 =59 .25 ,P<0 .000 1) .There was no significant difference of IgM positive rate between the planned pregnancy women who had closed contact to cats and the control group (χ2 =0 .23 ,P=0 .628 6) ,while the positive rate of IgG was higher than that of the control group (χ2 =9 .95 ,P=0 .001 6) .T .gondii infec‐tion rate was on the low level of planned pregnant women in Chongqing .Adverse pregnancy outcomes are related to Toxoplas‐ma infection .
5.Investigation on distribution of Yersinia enterocolitica in Dengfeng f rom 2009 to 2012
Hongmin LI ; Jiayong ZHAO ; Shengyong ZHANG ; Lili HUANG ; Dexiang WANG ; Changmin ZHAO ; Shengli XIA
Chinese Journal of Zoonoses 2015;(3):281-283
To investigate the infection of Yersinia enterocolitica in Dengfeng City ,the strains were isolated from livestock and poultry .The strains were detected with biochemiological methods ,serological methods ,and virulence genes were detected with PCR .A total of 105 strains of Yersinia enterocolitica were classified from 1 285 stool samples ,the total isolation rate was 8 .17% .Among the total isolated strains ,17 strains were classified from dogs with a rate of 17 .35% and 35 strains from pigs with 13 .62% .Twelve strains were O ∶3 serotype (13 .48% ) ,12 strains were O ∶5(13 .48% ) ,and 14 strains were O ∶8 (15 .73% ) .Ail+ ,ystA+ ,yadA+ and virF+ accounted for 12 .36% ,and ystB+ accounted for 42 .70% .In conclusion ,the pigs and dogs were important animal hosts ,which may play the major role in humans'infection .
6.Application of spatial statistics on echinococcosis studies
Chinese Journal of Zoonoses 2015;(3):272-276
Spatial statistics plays an important role in spatial epidemiology studies of echinococcosis .Spatial statistics can be used to describe the spatial distribution ,predict the prevalence ,identify disease clusters ,and analyze the influencing factors of echinococcosis .To describe spatial distribution and predict the prevalence ,we can use spatial interpolation ,empirical bayes smoothing and ellipsoidal gradient .Spatial autocorrelation always used to identify disease clusters .Moran's I value ,Getis'G val‐ue and spatial scan statistics are used to judge spatial autocorrelation .Molding plays an important role on analyzing risk factors of echinococcosis .Generalised linear mixed models and Bayesian model are always performed with both spatial factors ,such as geomorphologic features ,climatic characteristics ,vegetation index and factors which make great effect on disease transmission . To figure out the spatial distribution of echinococcosis is significant for echinococcosis control and prevention .
7.Problems and challenges in prevention and control of animal hydatid disease
Xingju SONG ; Dandan HU ; Guangyou YANG
Chinese Journal of Zoonoses 2015;(3):264-271
Hydatid disease does seriously harm to human and livestock ,and causes huge economic losses to the livestock in‐dustry .Despite the fact that people have made some success in prevention and control of animal hydatid disease after making great efforts during the past few decades ,however ,there still remain many problems and challenges .In order to facilitate the research in animal hydatid disease in China ,here we reviewed the problems and challenges in the prevention and control of this disease and put forward several proposals on the treatment and management of dogs ,immunization ,diagnose ,surveillance , etc .
8.KPC carbapenemases among Enterobacteriaceae with carbapenem non-susceptibility
Hui HUANG ; Shumin SHE ; Ximei ZHAN ; Duorong WU
Chinese Journal of Zoonoses 2015;(3):247-250
To investigate the prevalence and gene types of KPC in Enterobacteriaceae strains isolated from 4 tertiary general hospitals in Hainan area ,a total 43 isolates which were resistant or intermediate to imipenem or ertapenem were collected from sterile sites between August 2012 and June 2013 from 4 tertiary general hospitals in Hainan area .Modified Hodge Tests (M HT ) were performed for KPC phenotype screening .PCR amplification and DNA sequence were performed to analyze the encoding genes of KPC .Results showed that in the 43 isolates ,21 strains were positive in M HT .PCR and DNA sequence analysis confirmed that 3 isolates produced KPC‐2 .It's suggested that there were the Enterobacteriaceae carrying KPC in Hain‐an area .The encoding genes were KPC‐2 .The KPC gene could be horizontally transmitted by plasmid among different groups of bacteria .It is important to control the transmission of these Enterobacteriaceae carrying KPC .
9.Virulence gene detection and multi-locus sequence typing of Vibrio parahaemolyticus from patients in Ningbo,China
Hong GAO ; Qifa SONG ; Jingye XU ; Jian ZHENG ; Xuanyi SHEN
Chinese Journal of Zoonoses 2015;(3):240-243
To investigated the toxin genes distribution and molecular characteristics of Vibrio parahaemolyticus from pa‐tients in Ningbo ,V .parahaemolyticus strains were collected from patients with food poisoning and diarrhea .Thermostable di‐rect hemolysin gene (tdh) and TDH‐related hemolysin gene (trh) were detected by polymerase chain reaction (PCR) .Molecu‐lar characteristics were acquired by multi‐locus sequence typing (MLST ) .Of 248 clinical strains were isolated from 2006 to 2012 .Forty‐eight strains were selected to detect virulence genes and MLST genotyping .Forty‐two isolates were detected as tdh+ and 11 isolates were detected as trh+ .There were 9 STs and one undifferentiated type in Ningbo clinical strains .Thirty‐two strains were classified into ST3 ,5 strains into ST265 and 3 strains into ST120 .ST265 was found in Ningbo strains com‐pared with strains from other regions of China .Strains with tdh+ accounted for the majority in Ningbo clinical strains .Twen‐ty‐five strains of ST3 clone were tdh+ /trh‐.There were 9 STs coexsited in Ningbo clinical strains .ST3 clone was dominant , followed by ST265 and ST120 .Strains with tdh+ /trh‐were dominant in the ST3 clone .The unique ST262 was found in Ning‐bo clinical strains .
10.Detection and phylogenetic analysis of arenavi rus carried by wild rodents in Ningbo,China
Qun HU ; Sijie MA ; Chunyin ZHOU ; Shumei TONG ; Yong MEI
Chinese Journal of Zoonoses 2015;(3):235-239
To detect and phylogeneticaly analyze arenavirus carried by wild rodents in Ningbo ,China ,two pairs of degener‐ate‐primers were designed to amplify the S and L gene of arenavirus ,and then RT‐PCR was applied to detect arenavirus carried by rodents which captured from Ningbo port area .All 73 rodents samples were detected ,of which 12 Rattus norvegicus were positive ,an arenavirus virus strain named DX1401 were separated .The S gene amplified products of DX1401 was about 413 bp ,and the L gene was 1 204 bp .The phylogenetic analysis of S segments showed that DX1401 strain was in one branch of phylogenetic tree with Mobala virus strain ACAR3080 .The genetic distance to Mobala virus strain ACAR3080 was the closest , with the value of 0 .467 ;the phylogenetic analysis of L segments showed that DX1401 strain were in one group of phylogenetic tree with Lassa virus strain Josiah ,NL ,Z148 ,Bamba‐R114 ,Soromba‐R ,Nig08‐A37 ,Nig08‐A47 ,Mobala virus strain ACAR3080 ,Morogoro virus strain 13017/2004 ,Mopeia virus strain Mozambique ,and AN 21366‐BNI .The genetic distance to Mobala virus strain ACAR3080 was the closest ,with the value of 6 .953 .In conclusion ,the study confirmed the existence of arenavirus popular in wild rodents in Ningbo ,China .