The mutation characteristics of drug-resistance of Mycobacterium tuberculosis in Qinzhou,Guangxi / 国际检验医学杂志

Objective To analyze the variation characteristics of rpoB,katG,inhA,rpsL and embB related genes of Mycobacterium tuberculosis(MTB)in Qinzhou,Guangxi.Methods PCR reverse point hybridization was used to detect 5 common resistance mutants of Mycobacterium tuberculosis in 237 MTB-DNA positive sputum samples.Results Among 237 cases of tuberculosis patients,72 cases(30.38%)were resistant to the four kinds of anti-TB drugs.The resistance mutation rate of rifampin,isoniazid and streptomycin was 2.53%, 13.92%,3.80%.The top 5 gene mutation detection loci of Mycobacterium tuberculosis were-15M,S531L and 43M.Conclusion The main drug-resistant strains are isoniazid resistance,and the mutation of inhA gene were the major one in Qinzhou,Guangxi.

Application of PCR－reverse dot blot hybridization gene membrane chip technology in genetic testing of children with hereditary nonsyndrome deafness / 中华实用儿科临床杂志

Objective To explore the application value of PCR－reverse dot blot hybridization (PCR－RDB) gene membrane chip technique in genetic diagnosis of hereditary non－syndrome deafness in children.Methods The blood samples(2 mL)were collected from 38 children with congenital deafness,excluding high－risk factors for deaf-ness at Dongguan Rehabilitation School,and then genomic DNA extracted.By using self－designed multiplex－PCR combined with PCR－RDB gene chip technology,20 hot－spot mutations of 4 pathogenic genes of common deafness in Chinese population were detected.Sanger sequencing was used as the gold standard to corroborate the positive samples. Results Among 38 subjects,deafness gene mutations were detected in 16 cases,with a detection rate of 42.11%,and they were all verified by family study.Among 16 cases,6 cases of GJB2 gene mutation(3 cases of homozygote,3 cases of heterozygous),4 cases of SLC26A4 mutation,2 cases of MTRNR (m.1555A＞G)mutation,4 cases of compound muta-tion,but none of GJB3 gene mutations.And their detection rates were 15.79%,10.53%,5.26%,10.53%,and 0,re-spectively.DNA samples from 16 children with deafness gene mutation were corroborated by Sanger sequencing,and the compliance rate was 100%.Conclusions For 20 hot－spot mutations of 4 common deafness pathogenic genes,the matc-hing PCR－RDB gene membroine chip technology was designed and the susceptible gene of congenital deafness children was detected.This technique has some advantages like high detection rate,fast,accurate and economical.It is an ideal method for gene screening on hereditary non－syndrome deafness children and has good clinical application prospects.

Application of PCR-reverse dot blot hybridization in the diagnosis of vulvovaginal candidiasis and the detection of Candida albicans drug-resistant genes / 临床检验杂志

Objective To evaluate the application value of PCR-reverse dot blot hybridization in the identification of Candida and the detection of Candida albicans drug-resistant genes.Methods The vaginal secretion samples from 285 patients with candidal vaginitis and 50 healthy women were collected.The identification of Candida species and their drug susceptibility were detected by the bioMérieux Yeast identification cards and MIC method(Zhengzhou Antu kit),respectively.The identification of Candida species and the mutation of Candida albicans,drug-resistant genes were also detected by the Shenzheng Yaneng test kit(PCR-reverse dot blot hybridization).The drug-resistant genes were also identified by PCR and nucleic acid sequencing.Based on the culture identification,MIC method and nucleic acid sequencing as the contrast methods,the sensitivity,specificity and accuracy of PCR-reverse dot blot hybridization in the identification of Candida species and the mutation detection of Candida albicans drug-resistant genes were evaluated.Results Compared with the bioMérieux Yeast identification method,the sensitivity,specificity,positive predictive value,negative predictive value and total coincidence rate of PCR-reverse dot blot hybridization for detecting six kinds of Candida species,including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilosis,Candida krusei and Candida guilliermondii,were above 95％,96％,96％,98％ and 97％,respectively.There was no significant difference in detecting six kinds of Candida species between the two methods (x2 =0.44,0,0,0,0 and 0,respectively,P ＞ 0.05),and there was good consistency between them (Kappa ＞ 0.9).Compared with the MIC method,the sensitivity,specificity,positive predictive value,negative predictive value and total coincidence rate of PCR-reverse dot blot hybridization for detecting the drug resistance of Candida albicans were 98％,88％,98％,88％ and 96％,respectively.There was no significant difference in detecting the drug resistance of Candida albicans between the two methods (x2 =0.17,P ＞ 0.05),and there was good consistency between them (Kappa ＞ 0.8).The results of PCR-reverse dot blot hybridization in detecting the mutation sites of six kinds of Candida albicans drug-resistant genes were 100％ of coincidence with that of the nucleic acid sequencing method.Conclusion The PCR-reverse dot blot hybridization has high consistency with the culture method and the nucleic acid sequencing method in the identification of Candida species and the mutation detection of Candida albicans drug-resistant genes,which is more early and rapid than the traditional detection methods,and may be applied to the auxiliary diagnosis of vulvovaginal candidiasis (VVC).

Analysis of the distribution of various types of HPV and their influencing factors among women with normal cervical cytology in the Shanghai area / 中华健康管理学杂志

Objective To understand the various HPV types and the factors influencing their distribution among women with normal cervical cytology in the Shanghai area, to provide basic data for cervical cancer prevention and vaccine use.Methods A questionnaire-based survey was conducted among 3 372 married women in the Shanghai area. A PCR reverse dot blot (PCR-RDB) method was adopted for HPV genotyping of cervical exfoliated cell samples from 3 206 women. Odds ratio (OR) for HPV infection were analyzed using logistic regression. Results Of the 3 206 women investigated, 669 (20.87%) were positive for HPV infection. The highest incidence of HPV infections was seen in the age groups of 55-59 years, 50-54 years and 45-49 years, with the rates of positive detection being 27.67%, 21.65%, and 21.55%, respectively. While 70.4% of the positive cases had a single infection, 29.6% showed multiple infections. In cases with multiple infections, double infection was predominant (20.63%). The top five high-risk gene types were HPV52(3.65%),HPV53(2.71%),HPV51(2.03%),HPV58(1.87%),and HPV16 (1.40%). The top three low-risk gene types were HPV81 (2.03%), HPV42 (1.43%), and HPV55 (1.31%). Among the women with HPV infections,there were 483 having medium-high-risk HPV infection.There were significant differences in the age,the age at first sexual activity,sexual activities per week,education,and alcohol consumption between HPV positive and HPV negative patients. Age stratification showed that the infection rates in the 55-59 years old group were significantly higher than that in the other age groups(χ2=15.349, P=0.000). Both single factor and multivariate non-conditional logistic regression analyses showed that higher education and the start of sexual activity at a later age were protective factors for medium-high-risk HPV infection,with regression coefficients of-0.165 and-0.08,respectively (P<0.01) in the multivariate analysis. The risk factors included age between 55- 59 years, menstrual status (menopause), sexual activity (≥3 times per week) and alcohol consumption. High-risk HPV infections also had the same risk factors,and the odds ratios were 1.558,1.275,and 1.678,respectively(P<0.01).However, 55-59 years of age and alcohol consumption are independent risk factors for medium-high risk HPV infection. Conclusions High-risk HPV in women of Shanghai is commonly caused by HPV52, HPV53, HPV51,HPV58,and HPV16.The high-risk group includes women who are 55-59 years old.While drinking is an independent risk factor for medium-high risk HPV infection, a moderate sex life and delayed age at first sex can reduce the risk of high-risk HPV infections.

Verification application of PCR techniques with different principles in prenatal diagnosis of thalassemia / 实用医学杂志

Objective To evaluate the necessity and feasibility by using two different PCR-based techniques for prenatal diagnosis of thalassemia. Methods 509 specimens for prenatal diagnosis of thalassemia were detected respectively by single tube multiplex PCR(STMP),reverse dot blot(RDB)or probe melting curve assay(PMCA)for commonα-thalassemia orβ-thalassemia mutations in double-blind tests. Samples with different detection results were confirmed with DNA sequencing analysis. Prenatal diagnosis of thalassemia results were verified or followed up after birth. Results In detectingα-thalassemia andβ-thalassemia,there was one case in STMP + RDB and another case PMCA indicating differentiating results. The detection sensitivity of STMP + RDB was higher than that of PMCA,and its difference can be used as an indication for maternal blood contamination. Conclusion The two PCR methods with different principles were necessary and feasible for the prenatal diagnosis of thalassemia. The two methods were complementary to each other ,which can ensure the reliability of the prenatal diagnosis results and reduce the defects of single technique. It is worthy to be popularized in clinical application.

Diagnostic value of different detection methods for Mycobacterium tuberculosis in bronchoalveolar lavage fluid from pulmonary tuberculosis patients / 中南大学学报(医学版)

Objective:To investigate the diagnostic value of different detection methods for Mycobacterium tuberculosis in bronchoalveolar lavage fluid (BALF) from patients with pulmonary tuberculosis.Methods:BALF from l00 patients in Changsha Central Hospital from January 2013 to December 2015 was collected.Among 100 patients,65 cases were clinically diagnosed as tuberculosis,and 35 cases served as control.BALF smear method,polymerase chain reaction (PCR) and membrane reverse dot blot (RDB) were used for synchronous detection of Mycobacterium tuberculosis.Results:The positive rates by BALF smear method,PCR and RDB were 43.08％,73.84％ and 92.31％,respectively (P＜0.05).Sensitivity,specificity,accuracy,and negative predictive value for BALF smear were 43.08％,88.57％,59.00％,and 45.59％,respectively;for PCR were 73.85％,100％,83.00％,and 67.31％,respectively;for RDB were 92.31％,100.00％,95.00％,and 87.50％,respectively.Conclusion:The technique of membrane RDB can not only accurately diagnose Mycobacterium tuberculosis,but also can rapidly and easily identify the resistance of Mycobacterium tuberculosis to streptomycin (SM),rifampicin (RFP) and isoniazid (INH) genotypes.It possesses high clinical value.

Analytical research on HPV infection among women in Xingjiang Kashgar area / 国际检验医学杂志

Objective To investigate the exfoliative cell HPV infection situation,genotype distibution and charactristics among female population in Kashgar area.Methods The cervical exfoliated cells specimens were collected from 1548 women,and 23 genotypes of HPV were detected by using the PCR reverse dot blot hybridization method.The HPV infection situation and its genotypes were analyzed.Results The HPV infection positive rate was 33.33% (485/1 548),in which the HPV-16 infection rate was 10.3%;HPV52 infection rate was 9.9%;HPV58 infection rate was 7.6%,HPV53 infection rate was 7.2%;the single subtype infection rate was was 66.8%,the multiple infection rate was 33.2%;the HPV infection rate was highest(43.3%) in female population aged 41-50 years old,which was extremely higher than that in other age groups(P<0.05).Conclusion The HPV infection among female population of Kashgar area is dominated by HPV16,the HPV infection rate is highest in female population aged aged 41-50 years old,which provides a theoretical basis for the HPV prevention and control work of related departments in Kashgar area.

Analysis of 1 817 cases of papilloma virus infection in women / 国际检验医学杂志

Objective To investigate the infection status and genotype distribution of human papillomavirus (HPV) infection in Changzhou district ,and to provide a theoretical basis for the prevention ,development and clinical diagnosis and treatment of HPV . Methods From October 2015 to July 2016 ,1 718 cases of female cervical epithelial cells were collected ,and 28 kinds of gene typing were detected by PCR‐reverse dot blot hybridization .Results The infection rate:1 718 cases of women were collected ,the positive HPV infections were 34 .23% .The infection types :single infection rate was 23 .57% (405/1 718) .The high‐risk HPV subtype in‐fections accounted for 17 .17% (295/1 718) and the low‐risk HPV subtype infections accounted for 5 .18% (89/1 718) ,suspected high‐risk infection was 1 .22% (21/1 718) .Multiple infection rate was 10 .94% (188/1 718) .HPV52 was the most common infec‐tion among high‐risk HPV infection ,the positive rate was 16 .16% (95/588) .HPV61 was the most common infection among low‐risk HPV infection ,the positive rate was 4 .08% (24/588) .There was no significant difference between age and HPV positive rate . The 61-70 age group had the highest HPV multiple infection rate in all age groups .Conclusion The high HPV infection is ob‐served in Changzhou district ,among which single HPV52 infection and the high‐risk HPV infection are the most common infec‐tions .There is difference in HPV infection among different age groups .

Analysis of HPV subtype infection in 4 037 healthy women in Haikou area / 中华微生物学和免疫学杂志

Objective To investigate the infection of 23 kinds of human papillomavirus ( HPV) subtypes in female cervical epithelium samples and to analyze their relationships with age and results of cyto-logic test in Haikou area. Methods A total of 4 037 local healthy women were enrolled in this study from July 2013 to August 2016, 1 967 of whom received cervical cytology test. Cervical cell samples collected from those women were detected for HPV typing by using PCR-reverse dot blot hybridization. Results (1) The total positive rate of HPV in 4 037 samples was 22. 15% (894 cases), and the detection rates of carci-nogenic, possibly carcinogenic and non-carcinogenic HPV were 16. 13%, 3. 99% and 5. 55% (651, 161 and 224 cases), respectively. The positive rates of 6 genotypes were high, which were HPV52, 53, 81, 51, 16 and 58 in turn. (2) The detection rates of carcinogenic, possibly carcinogenic HPV and some of the gen-otypes (HPV18, 52, 53, 66) increased with age (all P<0. 05). (3) Multiple infection in HPV-positive women accounted for 24. 38% (218/894), in which the infection rates of carcinogenic types declined with age and the numbers of HPV genotypes in carcinogenic infections were negatively correlated with age ( both P<0. 05). (4) Only 2. 49% (49/1 967) of the samples were positive for cervical cytologic test, classified into the ≥ASC-US ( atypical squamous cells of undetermined significance ) group. The detection rates of eight kinds of carcinogenic and two kinds of possibly carcinogenic HPV in≥ASC-US group were significantly higher than those in the negative (no intraepithelial lesion or malignant lesion, NILM) group (all P<0. 05). Conclusion This study indicates that Haikou women have higher rates of HPV infection, especially the eld-erly group. HPV subtype infections present some regional characteristics and are mainly single type infec-tion. Cervical cancer screening should be combined with tests for HPV and cytology analysis to improve its effectiveness.

The status of human papillomavirus infection in one area / 国际检验医学杂志

Objective To analyze the genotypes and age distributions of human papillomavirus in affiliated hospital of Chifeng u‐niversity ,to provide theory basis for the screening and prevention of cervical cancer in Chifeng area .Methods The cervical exfolia‐ted cell specimens from 1 368 gynecological outpatients and female inpatients in affiliated hospital of Chifeng university were collect‐ed from January to June 2016 .Using PCR‐reverse dot blot technology ,25 HPV subtypes was performed ,a statistical analysis was conduted combing with age .Results Among 1 368 specimens ,546 specimens were found with positive HPV ,the positive rate was 39 .91% .The 24 genotypes were detected .The top three subtypes of HPV infection were HPV16(12 .08% ) ,HPV 58(8 .05% )and HPV 52(7 .61% ) ,HPV73 genotype was not detected .Masculine gender rate in the groups with various age had significance differ‐ence after the chi‐square test ,among different age groups ,the prevalence rate was the highest in the patients with higher than 56 years old(52 .5% ) ,followed by the rate in patients with lower than 25 years old(52 .38% ) .Conclusion The subtype‐specific and age‐specific distributions of HPV infection in Chifeng area have obvious heterogeneity ,which indicates that HPV genotypes and age should be considered in screening ,prevention and treatment of cervical cancer .

Study on relationship between hepatitis B virus genotypes and drug resistance mutation / 国际检验医学杂志

Objective To explore correlations between hepatitis B virus (HBV)genotyping,other clinical information and drug resistance mutations.Methods 358 cases of patients with chronic hepatitis B (CHB)were selected as subjects,and the resistance loci and genotypes were detected by using the polymerase chain reaction(PCR)-reverse dot blot technique.Clinical data,such as ser-um HBV DNA loads,serum levels of alanine aminotransferase(ALT)and hepatitis B virus e antigen(HBeAg),gender,age and length of nucleoside analogues use were collected.Results All samples were successfully amplified positive band.Type B(267 ca-ses)was the main HBV genotype,followed by type C(81 cases)and type D(10 cases).In the 31 1 cases of patients taking nucleoside analogues,269 cases were completely wild type.No drug resistance mutation was found in 47 cases of patients not taking medicine. The drug resistance mutations mainly occured in 204 and 180/204 site.There was no significant correlation between resistance mu-tations and gender,age,serum HBV DNA loads,genotype,serum levels of ALT and HBeAg(P >0.05).While the medication time was longer,the incidence of resistant mutants was greater(P <0.05).The 180 mutation had a certain correlation with 204 site mu-tation(P <0.05).Conclusion PCR-reverse dot blot technology can effectively detect the HBV genotype and mutations,which could effectively guide the clinical medication.

Research of hepatitis B virus genotyping and resistance mutation gene detection / 国际检验医学杂志

Objective To investigate the distribution characteristics of hepatitis B virusgene types and the situation of drug re-sistance gene in Chuxiong area and to benefit the rational clinical antiviral therapy and the prognostic evaluation.Methods The DNA content,hepatitis B virus gene types and drug resistance associated sites in serum of hepatitis B patients were detected by the technology of fluorescence quantitative PCR and the gene chip of reverse dot blot.Results Among 103 serum samples,detection of DNA content in 85 samples was positive while DNA content of 18 samples was negative.85 positive samples were typed for hepati-tis B virus DNA,of which 63 samples were classified as type C(74.1%),21 samples were classified as type B(24.7%)and 1 sample was classified as mixed type B and C (1.2%).57 samples had the wild-type gene loci(67%)and 28 samples had the mutant gene locus(33%).Among these samples,1 7 cases of mutation were placed in rt180M,12 cases were placed in rt204V,6 cases were placed in rt204I,3 cases were placed in rt207I,3 cases were placed in rt181V and 3 cases were placed in rt236T.Among 28 samples of drug resistance gene mutation,5,22,24 and 1 1 samples were sensitive to lamivudine,telbivudine,adefovir and Entecavi,respectively.Con-clusion The main hepatitis B virus genotype is type C in Chuxiong area.The most important mutant site is rt180M and the second is rt204V.The most sensitive drugs are telbivudine and adefovir in clinical antiviral therapy and the second sensitive drug is Ente-cavi While the most commonly resistant drug is lamivudine.

Rapid detection of rifampin-resistant clinical isolates of Mycobacterium tuberculosis by reverse dot blot hybridization / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in 'hot mutation region' of Mycobacterium tuberculosis (M. tuberculosis).</p><p><b>METHODS</b>12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay.</p><p><b>RESULTS</b>The sensitivity and specificity of the RDBH assay were 91.2% (165/181) and 98.3% (117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7% (293/300), 98.2% (164/167), and 97.0% (129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively.</p><p><b>CONCLUSION</b>Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.</p>

Survey and research of common thalassemia genotypes in northeast area of Chongqing / 国际检验医学杂志

Objective To understand the gene carrying rate ,gene mutation types and distribution characteristics of thalassemia in the northeast area of Chongqing .Methods 28 633 specimens collected from the patients and individuals with physical examina‐tion in our hospital from January to December 2013 were performed the RBC parameters detection and hemoglobin electrophoresis screening .The specimens with phenotype positive were definitely verified the thalassemia type by using Gap‐PCR and reverse dot blot(RDB) .Results Among 28 633 specimens ,1 358 specimens were finally diagnosed as thalassemia with the thalassemia carrying rate of 4 .74% ,including 589 cases(2 .06% ) of α‐thalassemia and 741 cases (2 .59% ) of β‐thalassemia cases .Among the α‐thalasse‐mia genotypes ,‐αα/‐‐SEA genotype(1 .38% ) was most common ,the next was ‐αα/‐α3 .7 genotype (0 .37% ) and αα/‐α4 .2 genotype (0 .20% ) .Among the β‐thalassemia genotypes ,CD41‐42 genotype (1 .27% ) had the highest constituent ratio ,followed by IVS‐2‐654 genotype(1 .27% ) and CD17 genotype(0 .30% ) .28 cases were found to be the double heterozygote with α‐thalassemia and β‐thalassemia .Conclusion The northeast area of Chongqing is a region with the high incidence rate of thalassemia and complicated heredity .Therefore this research provides the reference information for the prevention of thalassemia ,genetic counseling and prena‐tal diagnosis .

Analysis of women′s genital human papillomavirus infection and genotype distribution in Shenzhen / 国际检验医学杂志

Objective To investigate the female human papillomavirus(HPV)infection situation in Baoan district,the HPV pos-itive rates in different age groups and the subtypes distribution.Methods PCR followed with reverse dot blot was performed to ex-amine 23 kinds of HPV genotypes in 2 627 female patients in our hospital from the January 2011 to December 2012.Results In 2 627 samples,the positive rate of HPV was 23.94% (629 cases),in which the infection rate of single low-risk type was 15.1%(95 cases),the main HPV genotype was HPV43 (7.79%);the infection rate of high-risk type was 55.17% (347 cases),the 3 most prevalent HPV genotypes were HPV52 (12.56%),HPV16 (9.86%)and HPV58 (7.79%).The multiple HPV infection ac-counted for 29.73% (187 cases).The HPV infection rates in different age groups were 50.0% in age 15-20 years,24.7% in age 21-30 years,20.8% in age 31-40 years,25.8% in age 41 -50 years,42.1% in age >50 years respectively,the differences had statistical significance.Conclusion The HPV infection rate is 23.94% in Bao′an district.The most prevalent HPV genotypes are HPV 52,16,58,43.Women in age 15-20 years old have a higher infection rate.

Application of reverse dot blotting for detection of Tyzzer’ s organism / 中国实验动物学报

Objective To establish a simple, stable, specific and sensitive method for detection of Tyzzer ’ s or-ganism by reverse dot blotting ( RDB) .Methods Primers and specific probes were designed according to the conservative sequence of Tyzzer 16S rDNA.The forward primer was labeled with biotin .The reverse dot blotting method was established followed by PCR amplification .The specificity and sensitivity of this method were determined .Next, 41 mice and 38 rats were examined by RDB , ELISA and IFA .Results The RDB method showed a high specificity , and in the testing of the 79 laboratory animals , its limit of detection was 4.5 ng/μL.Compared the results of ELISA and IFA , its consistence with ELISA was 100%and the positive rate was 7.59%(6/79), the consistence with IFA was 92.4%(73/79), and the posi-tive rate was 0%.Conclusions An accurate, sensitive and specific method in combination with PCR and RDB in detection of Tyzzer’ s organism is established in this study .

Correlation analysis of positive rate of HPV genotyping test and HPV nucleic acid loads / 中华微生物学和免疫学杂志

Objective To investigate the correlation between the positive rate of PCR-reverse dot blot genotyping test and the loads of the viral nucleic acid. Methods The fluorescent PCR assay was used to detect the high-risk HPV(HR-HPV)DNA loads in the cervical mucus samples from 1162 female pa-tients. Patients with positive HR-HPV DNA were further analyzed by PCR-reverse dot blot hybridization as-say for HPV genotyping. Results The overall positive rate of genotyping test was 68. 8% . The positive rate of genotyping test had a significant positive correlation with the Log Koc of HR-HPV DNA loads(r=0. 944, P﹤0. 01). The quadratic curve fitting formula was Y= -1. 806+0. 558X-0. 031X2(Y for genotyping positive rate,X for Log Koc of HR-HPV DNA loads). There were significant differences with the positive rate of gen-otyping test among patients with different viral loads(P﹤0. 01). When HR-HPV DNA loads were 104-105 copies/ ml,105-106 copies/ ml,106-107 copies/ ml and more than 107 copies/ ml,the positive rate of HPV genotyping test were 27. 8% ,48. 5% ,74. 0% ,97. 5% and 33. 3% ,51. 5% ,78. 0% ,97. 5% respective-ly by using different genotyping detection reagents. Conclusion The positive rate of PCR-reverse dot blot genotyping test was correlated with the loads of HPV nucleic acid.

Infection situation and type distribution characteristics of human papillomavirus in Fengxian District of Shanghai / 国际检验医学杂志

Objective To investigate the infection situation and the gene subtype distribution characteristics of human papilloma-virus(HPV)in Fengxian District of Shanghai to provide the reliable scientific basis for preventing HPV infection and the prevention and treatment of cervical cancer.Methods The PCR combined with reverse dot-blot technique was adopted to analyze the HPV de-tection results among 18 194 women in our hospital from February 2011 to October 2013.Results Among 18 194 cases of sample, 2 986 cases were HPV positive with the total HPV infection rate of 16.41%,in which 21 genotypes were all detected out.Single HPV genotype infection was in 2714 cases,two kinds of HPV genotype infection were in 148 cases(4.96%)and three kinds or more of HPV genotype infection were in 124 cases.The HPV positive rate was 16.88% in the outpatient and 12.76% in the inpa-tients.In the HPV infection,the high-risk type was more than the low-risk type,its detection number was 7.5 times of low-risk type.Most of the high-risk HPV infection were HPV16,52,58;most of the low-risk HPV infection was HPV cp8304,followed by HPV6 and HPV11.Conclusion The HPV prevalence in Fengxian District is generally in line with the Asian population distribution rule,but has its own unique regional characteristics.The infection age peak is 51-65 years old.The high-risk type infection is high-er than the low-risk type infection,which is dominated by the single genotype infection.

Comparison of real time PCR and PCRˉreverse dot blot hybridization for detection of Human papillomavirus / 国际检验医学杂志

Objective To compare real time PCR with PCR-reverse dot blot hybridization (PCR-RDB)for detecting human pap-illomavirus (HPV)infection in women.Methods A total of 109 genital specimens from women were collected in the study.All specimens were tested HPV by using real time PCR and PCR-RDB,discrepant samples were tested again by PCR-xMAP.Results The concordant rate was 83.5%(91/109)between real time PCR and PCR-RDB (kappa=0.671),the other 18 discrepant samples were retested by PCR-xMAP,7 of those were identical with real time PCR and 11 with PCR-RDB.No differences of PCR-RDB pos-itive rates were found between the high and low viral load groups (χ2 =1.476,P =0.224).Conclusion It demonstrated moderate consistency between real time PCR and PCR-RDB.The HPV positive rates of PCR-RDB were stable when the viral loads were 103-108 .

Analysis of β-thalassemia mutations in Fujian province / 中国地方病学杂志

ObjectiveTo explore the prevalence and spectrum of β-thalassemia mutations in Fujian province,and to provide a reference for prenatal diagnosis and genetic counseling in this population.Methods Two thousand three hundred and one blood samples were randomly selected from 9 different areas of Fujian province from May 2008 to December 2010.PCR and reverse dot blot hybridization (RDB) were adopted for detection of the 17 common types of mutation,and the frequency of each genotype of β-thalassemia mutations was calculated.The β-globin gene of unknown positive samples were analyzed directly with DNA sequencing.Results Three hundred and fifty-nine cases were detected with β-thalassemia mutations of the 2301 copy blood samples submitted,and the detection rate was 15.60％ (359/2301).Of the mutated genes,12 different mutations were identified,namely IVS-2-654(C→T),CD41-42(-TCTT),CD17(A→T),-28(A→G),CD27-28(+C),CD26(G→A),CD71-72(+A),IVS-1-1(G→T),CD43(G→T),-29(A→G),initiation codon ATG→AGG and CD36(-C).Mutation frequencies were 46.54％ (175/376),33.24％ (125/376),9.31％ (35/376),5.05％ (19/376),2.13％(8/376),1.33％(5/376),0.80％(3/376),0.27％(1/376),0.27％(1/376),0.27％(1/376),0.53％(2/376),and 0.27％(1/376),respectively.The most common mutations were IVS-2-654 (C→T) and CD41-42 (-TCTT),which accounted for 79.78％(300/376) of total genetic mutations.In addition,a novel β-globin gene mutation CD36 (-C) allele was detected for the first time,the deletion of a nucleotide C at code 36 within exon 2 lead to a frameshift mutation that could result in a premature termination at code 60.Conclusions β-thalassemia mutations in Fujian province are complex with significant genetic heterogeneity.We present for the first time the detection of a new β-thalassemia mutation in the population:CD36(-C),which provides valuable information for genetic counseling and prenatal diagnosis in Fujian province.