ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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).
ABSTRACT
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.