| Objective:?1?To evaluate the accuracy of Westmead?HbWS?band by using polyacrylamide gel chain electrophoresis?PAGE?,and to provide a new method for screening and diagnosing HbWS from the protein peptide level.?2?To investigate the HbWS genotype and hematology phenotypic characteristics among Baise population and compare hematology phenotypic differences of Thalassemia with other genotypes.Methods:?1?Experiment research: Cases aged between 2 and 60,taking gene examination of Thalassemia from 2015 to 2016,were selected.Hematological tests,Hb analysis and gene testing were gold standard for Thalassemia.At the same time PAGE method was applied to detect HbWS,among them,HbWS with following characteristics were positive cases:G? position zone with hyperchromatic effect,A? position without hyperchromatic effect and HbF with normalvalue,or others were negative cases.The sensitivity,specificity and other indexes of PAGE diagnostic method were calculated.?2?Clinical research: Carriers?patients?who diagnosed with different genotypes of HbWS from 2010 to 2016 in outpatient clinic were selected as research objects.According to specific inclusion criteria and exclusion criteria,carriers?patients?of each type of HbWS mutation were selected as observation group,and other kinds of Thalassemia gene carriers?patients?were selected and divided into several control subgroups.Quantitative hematology analysis for all samples was carried out by automatic analyzer and hemoglobin automatic analyzer.Gap-PCR was used to detect the deletion of ?-Thalassemia and reverse dot blot?RDB?assay for the detection of non deletion of ?-Thalassemiaand ?-Thalassemia.SPSS 19.0statistical software was applied to deal with data of different groups.Results:?1?600 cases were in accordance with the requirement,and PAGE diagnosis forthe achieved samples showed that 130 cases were positive,and 470 cases were negative in HbWS zone.According to the gold standard method,126 cases with HbWS gene mutation were detected,and 474 cases without HbWS gene mutation.The diagnostic accordance rate of HbWS by the PAGE diagnosis was 98.00%,sensitivity was 96.83%,specific was 98.31%,positive predictive value was 0.9385,negative predictive value was 0.9915,positive likelihood ratio was 57.3and negative likelihood ratio was 0.0249.ROC curve was used to analyze the area under HbWS curve of PAGE diagnosis,AUC was 0.942 and 95% confidence interval was?0.893,0.991?.?2?A total of 292 cases diagnosed with different genotype of HbWS were enrolled.genetic types were divided into eight categories,included 174 cases of HbWS heterozygote?? WS?/???,accounting for 59.59%,43 cases of HbWS combined with light ?-Thalassemia?accounting for 14.73%?,30 cases of HbWS combined with other static type double heterozygote of ?-Thalassemia?accounting for 10.27%?,28 cases of HbH-WS disease?accounting for 9.59%?,8 cases of containing HbWS type of light ?-Thalassemia combined with ?-Thalassemia?accounting for 2.74%?,4 cases of HbH-WS disease combined with light ?-Thalassemia?accounting for 1.37%?,3 cases of HbWS composite heavy ?-Thalassemia?accounting for 1.03%?and 2 cases of HbWS homozygote?accounting for0.68%?.?3?Hb?129.68±21.11g/L?,MCV?86.00±5.36fl?and MCH?27.76±2.16pg?of 117 cases in HbWS heterozygote group were all within normal range,the mean of them was lower than that of the normal group but higher than that of ?CS?/?? group,?QS?/?? group,??/-?3.7group and ??/-?4.2 group.RDW-CV?13.10 ±1.10%?and other static type ?-Thalassemia was no statistically significant difference but higher than that of the normal group.?4?Hb of double heterozygote in HbWS composite with other static type ?-Thalassemiawas within thenormal range?128.60±15.67 g/L?,higher than that of HbWS composite light ?-Thalassemia group and light ?-Thalassemia group.The MCV?80.27±4.70 fl?was within the critical range,and MCH?25.29±1.52 pg?was below the normal reference range,but higher than that of HbWS composite light ?-Thalassemia group.RDW-CV?13.38±1.27%?was lower than that of t the HbWScomposite light ?-Thalassemia group,light ?-Thalassemia group and the standard ?-Thalassemia.?5?The Hb?112.59± 14.03 g/L?,MCV?64.48±5.53 fl?and MCH?19.72 ± 1.81 pg?of the HbWS composite light ?-Thalassemia group were lower than the normal range,showing the cellule low pigment mild anaemia.and difference of Hb,MCV and MCH of light ?-Thalassemia group was not statistically significant.RDW-CV?15.67±1.34%?was lower than light ?-Thalassemia group.?6?The Hb?104.67±27.03 g/L?,MCV?67.76± 6.10 fl?,MCH?20.64 ±1.82 pg?of HbH-WS disease were the same as-?4.2/--SEAgroup and-?3.7 /--SEA group,showing cellule low pigmentmild anemia,but Hb level was higher than the latter two groups?P<0.05?.MCV of ? WS? /--SEA group was lower than ?CS? /--SEA group?P<0.05?.MCH had the least degree of diminution,while RDW-CV had the least degree of increase?P<0.01?..Conclusion:?1?The PAGE method that is HbWS zone positive,which the specificity,sensitivity and diagnostic coincidence rate of HbWS diagnosis are high,Accuracy approaches togenetic testing method,which can be used as HbWS new screening diagnosis methodFor the clinical application and Reference.In addition,the PAGE method can also detect the ?chain in the diagnosis of adult standard ?-Thalassemia.?2??WS?/?? is main genotype of HbWS in the Baise region.The hematology phenotype of ?WS?/?? is basically normal,and it is easy to be missed when screening.HbWS composite other static type ?-Thalassemia double heterozygote shows cellule low pigment.HbWS composite light ?-Thalassemia group shows cellule low pigment mild anemia,and HbH-WS disease also shows the cellule low pigment mild anemia,but anemia degree lighter than-?4.2/--SEA group?-?3.7/--SEA group.?3?Each ofthe HbWS different genotype hematology phenotype has its characteristic,but the degree of anemia is lighter than the same type Thalassemia.The study results have guiding significance for genetic counseling and prenatal diagnosis of thalassemia in the region. |
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