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.
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Objective To retrospectively analyze the result of HCMV‐DNA of breast milk and urine of neonatal hyperbilirubi‐nemia and evaluate the effect on screening neonatal hyperbilirubinemia .Methods Collected 461 cases of neonatal hyperbilirubinemia from January 2014 to December 2014 in our hospital ,which fed with breast milk as observation group ,at the same period collected 450 cases of healthy newborn in our hospital as control group .Tested the their level of HCMV‐DNA in breast milk and urine with Fluorescent Quantitative PCR ,recorded the testing results .Results 239 cases with positive HCMV‐DNA urine were detected in observation group ,accounting for 51 .8% .There were 89 cases of positive HCMV‐DNA urine in control group ,which accounted for 19 .8% ,the comparative difference was statistically significant(P< 0 .01) .There were 367 cases of positive HCMV‐DNA breast milk in observation group ,accounting for 79 .6% .The positive HCMV‐DNA breast milk were detected in 137 cases in control group ,which accounted for 30 .4% ,the comparative difference was statistically significant (P<0 .01) .Conclusion The breast milk of HCMV infection was an important way of neonatal HCMV infection ,the HCMV‐DNA detection of breast milk and urine was of great significance to the diagnosis of neonatal hyperbilirubinemia .
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Objective To retrospectively analyze the result of hemoglobin(Hb)test by using full-automatic Hb electrophoresis and evaluate the its significance in hemoglobinopathy.Methods The data of patients who underwent Hb electrophoresis test and regular blood tests in the hospital from January 2011 to December 2013 were included in the study.The test results were recorded including mean corpuscular volume(MCV),mean corpuscular hemoglobin(MCH)and results of Hb electrophoresis test.Final diag-nosis were made for suspected patients by using genetic testing,then disease detection rates and gene coincidence rates and constitu-ent ratios were calculated.Results 12 898 cases were included in the study,after statistical analysis the MCV was(85.32±13.61) fL,MCH was(29.87±6.44)pg.By using automatic hemoglobin electrophoresis,1 315 cases were found to be positive,in which 568 were male,747 were female,the detection rate was 10.19%.In the 1 315 patients,there were 761 cases suspectedα-thalassemia,ac-counted for 5.90%.There were 495 cases of suspectedβ-thalassemia,accounted for 3.84%,11 patients with HbJ(0.08%),15 pa-tients with HbK(0.12%),9 patients with HbG(0.07%),3 patients with HbD(0.02%),21 patients with HbE(0.16%).The sus-pected case′s final diagnosis were made by using genetic testing,α-thalassemia gene′s coincidence rate was 80.55%,β-thalassemia gene′s coincidence rate was 96.77%.Conclusion Automatic hemoglobin electrophoresis detection is of great significance for the di-agnosis of hemoglobinopathy.
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Objective To explore the genotype of patients with glucose-6-phosphate dehydrogenas(G6PD)deficiency in Dong-guan and provide the basis for the clinical diagnosis and prevention.Methods The clinical data of patients who took G6PD activity screening in the hospital were collected from January 2011 to December 2013,the G6PD/6PGD ratios were recorded.469 patients with positive G6PD/6PGD ratio were randomly enrolled in the study,whose mutations were detected by reverse dot blot(RDB)as-say.Results During this period,we measured G6PD activity of 16 464 cases by G6PD/6PGD ratios,there were 672 positive cases, the positive rate was 4.08%.Randomly selected 469 positive samples,detected their genotye by RDB assay.We detected 173 cases of G1376T,141 cases of G1388A,82 cases of A95G,60 cases of G871A,23 cases of G392T,14 cases of C1024T.In addition to that, we also found some rare mutations,such as 6 cases of C1004T,2 cases of T517C,1 cases of C1360T.65 cases of C1311T gene poly-morphism and 96 cases of dual gene mutations were detected.Conclusion The incidence of G6PD deficiency is high and the gene mutation types in Dongguan are both representative for Chinese population and with local heterogeneity.The study on gene muta-tions of G6PD deficiency is benefit for diagnosis and prevention.