廣西百色地區(qū)血紅蛋白Westmead的實驗及臨床研究
發(fā)布時間：2018-07-13 11:45
【摘要】：目的:(1)評價利用聚丙烯酰胺凝膠肽鏈電泳(PAGE)檢測異常血紅蛋白Westmead(HbWS)區(qū)帶的準(zhǔn)確性,從檢測蛋白質(zhì)肽鏈層面提供篩查和診斷HbWS的新方法。(2)探討百色地區(qū)人群中HbWS的基因型和血液學(xué)表型特點及其與其他不同基因型的地中海貧血(地貧)的血液學(xué)表型差異。方法:(1)實驗研究:收集2015年-2016年、籍貫系百色地區(qū)、在我院進(jìn)行地貧基因檢查、年齡在2-60歲之間的病例,以基因檢測作為診斷HbWS的金標(biāo)準(zhǔn)、同時應(yīng)用PAGE方法檢測HbWS區(qū)帶即具有Gγ位置區(qū)帶濃染而Aγ位置無濃染(HbF值正常)為HbWS區(qū)帶陽性,通過四格表計算PAGE陽性診斷HbWS的敏感性、特異性等各項指標(biāo)。(2)臨床研究:收集2010年-2016年、來源于百色地區(qū)、門診確診為HbWS的不同基因型攜帶者(患者)為研究對象,根據(jù)納入標(biāo)準(zhǔn)及排除標(biāo)準(zhǔn)從中篩選出HbWS各類型基因突變的攜帶者(患者)作為觀察組,篩選出其他相應(yīng)各類地貧基因型攜帶者(患者)分成若干對照組(亞組);所有樣本均進(jìn)行自動血細(xì)胞分析儀、血紅蛋白自動分析儀進(jìn)行血液學(xué)分析;采用Gap-PCR檢測缺失型α-地貧和反向斑點印跡(RDB)法檢測非缺失型α-地貧和β-地貧。應(yīng)用SPSS 19.0統(tǒng)計軟件對各組數(shù)據(jù)進(jìn)行處理。結(jié)果:(1)入組病例600例,取得的樣本經(jīng)PAGE實驗診斷HbWS區(qū)帶陽性130例、陰性470例;同時按金標(biāo)準(zhǔn)方法共檢出含HbWS基因突變的126例,無HbWS基因突變的474例。PAGE診斷HbWS的診斷符合率為98.00%、敏感度96.83%、特異度98.31%、陽性預(yù)測值0.9385、陰性預(yù)測值0.9915、陽性似然比57.3、陰性似然比0.0249。應(yīng)用ROC曲線分析PAGE診斷HbWS的曲線下面積AUC為0.942,95%置信區(qū)間為(0.893,0.991)。(2)收集確診為HbWS的不同基因型組合攜帶者(患者)共292例,基因類型組合分為八大種類,其中hbws雜合子(αwsα/αα)174例(占59.59%)、hbws復(fù)合輕型β-地貧43例(占14.73%)、hbws復(fù)合其它靜止型α-地貧的雙重雜合子30例(10.27%)、hbh-ws病28例(占9.59%)、含hbws的輕型α-地貧復(fù)合β-地貧8例(占2.74%)、hbh-ws病復(fù)合輕型β-地貧4例(占1.37%)、hbws復(fù)合重型β-地貧3例(占1.03%)和hbws純合子2例(占0.68%)。(3)117例hbws雜合子組的hb(129.68±21.11g/l)、mcv(86.00±5.36fl)、mch(27.76±2.16pg)在正常范圍內(nèi),其均值低于正常人組但高于αcsα/αα組、αqsα/αα組、αα/-α~(3.7)組和αα/-α~(4.2)組(該四個組mch均低于正常參考值);rdw-cv(13.10±1.10%)與其他靜止型α-地貧比較無差異但高于正常人組。(4)hbws復(fù)合其它靜止型α-地貧的雙重雜合子的hb在正常范圍(128.60±15.67g/l),高于hbws復(fù)合輕型β-地貧組和輕型β-地貧組;mcv(80.27±4.70fl)在臨界范圍、mch(25.29±1.52pg)低于正常參考值范圍,但高于hbws復(fù)合輕型β-地貧組;rdw-cv(13.38±1.27%)低于hbws復(fù)合輕型β-地貧組、輕型β-地貧組和標(biāo)準(zhǔn)型的α-地貧。(5)hbws復(fù)合輕型β-地貧組的hb(112.59±14.03g/l),mcv(64.48±5.53fl)、mch(19.72±1.81pg)低于正常范圍,表現(xiàn)為小細(xì)胞低色素性輕度貧血與輕型β-地貧組的hb、mcv和mch差異無統(tǒng)計學(xué)意義,rdw-cv(15.67±1.34%)低于輕型β-地貧組。(6)hbh-ws病的hb(104.67±27.03g/l)、mcv(67.76±6.10fl)、mch(20.64±1.82pg)與-α~(4.2)/--sea組、-α~(3.7)/--sea組一樣表現(xiàn)為小細(xì)胞低色素性輕度貧血但hb水平比后兩組高(p0.05);αwsα/--sea組的mcv低于αcsα/--sea組(p0.05),mch減低程度最輕,rdw-cv增加程度最輕(p0.01)。結(jié)論:(1)page方法hbws區(qū)帶陽性診斷hbws的特異性、敏感性和診斷符合率高,準(zhǔn)確性接近基因檢測方法,可作為hbws新的篩查診斷方法供臨床應(yīng)用參考;此外,page方法還可檢測ζ鏈診斷成年人標(biāo)準(zhǔn)型α-地貧。(2)百色地區(qū)hbws的基因突變類型主要以αwsα/αα為主,αwsα/αα的血液學(xué)表型基本正常,篩查時容易漏診;hbws復(fù)合其它靜止型α-地貧的雙重雜合子,表現(xiàn)為小細(xì)胞低色素;hbws復(fù)合輕型β-地貧組表現(xiàn)為小細(xì)胞低色素性輕度貧血;hbh-ws病也表現(xiàn)為小細(xì)胞低色素性輕度貧血,但貧血程度比-α~(4.2)/--SEA組、-α~(3.7)/--SEA組輕。(3)HbWS不同基因型血液學(xué)表型各有其特點,但是貧血程度都要比同型的地貧較輕,研究結(jié)果對該地區(qū)地貧的遺傳咨詢和產(chǎn)前診斷有指導(dǎo)意義。
[Abstract]:Objective: (1) to evaluate the accuracy of detection of abnormal hemoglobin Westmead (HbWS) zone by polyacrylamide gel electrophoresis (PAGE), and to provide a new method for screening and diagnosing HbWS from the detection of protein peptide chain. (2) to explore the genotype and hematological phenotypes of HbWS in the population of Baise and the Mediterranean with other different genotypes. Haematological phenotypic difference of anemia (ground poor). Methods: (1) experimental study: in 2015 -2016 years, the native place of Baise region, in our hospital in the poor gene examination, age between 2-60 years of age, gene detection as the gold standard for the diagnosis of HbWS, and the use of PAGE method to detect the HbWS zone with G gamma location with concentration and A gamma position no Strong staining (HbF value) was positive in HbWS area, and the sensitivity and specificity of PAGE positive diagnosis of HbWS were calculated by four grid tables. (2) clinical study: collect the different genotype carriers (patients) from the Baise area in 2010, from the clinic diagnosed as HbWS, and select the Hb according to the inclusion criteria and the exclusion criteria. The carriers (patients) of different types of gene mutations of WS were divided into several control groups (subgroups), and all the samples were carried out by automatic blood cell analyzer, hemoglobin automatic analyzer for hematological analysis, and Gap-PCR detection of deficient alpha and reverse dot print. The RDB method was used to detect the non missing alpha ground poverty and beta poverty. The data were processed with SPSS 19 statistical software. Results: (1) 600 cases were enrolled in the group, 130 cases of HbWS zone positive and 470 cases were diagnosed by PAGE test, 126 cases of HbWS gene mutation were detected by the gold standard method, and 474 cases of.PAG without HbWS gene mutation were found. The diagnostic coincidence rate of E diagnosis HbWS was 98%, sensitivity 96.83%, specificity 98.31%, positive predictive value 0.9385, negative predictive value 0.9915, positive likelihood ratio 57.3, negative likelihood ratio 0.0249. application ROC curve analysis PAGE diagnosis HbWS curve area AUC was 0.942,95% confidence interval (0.893,0.991). (2) collect confirmed HbWS different genotypes group A total of 292 patients (patients) were divided into eight types, including 174 hbws heterozygote (alpha WS alpha / ALA) (59.59%), 43 hbws complex beta poor ground poor (14.73%), 30 double heterozygotes (10.27%), hbh-ws disease 28 (9.59%), and 8 cases (2), hbh-ws disease, and 8 cases (2) (8 cases) with hbws. .74%), 4 cases of hbh-ws disease combined with light beta poor (1.37%), 3 cases of hbws complex beta poor (1.03%) and 2 cases of hbws homozygote (0.68%). (3) the HB (129.68 + 21.11g/l), MCV (86 + 5.36fl), MCH (27.76 + 2.16pg) in the hbws heterozygote group were in the normal range, which was lower than the normal group but higher than the alpha alpha / alpha alpha group, alpha alpha A A / A / alpha / A / - Alpha ~ (3.7) and alpha / - alpha ~ (4.2) groups (the four groups of MCH are lower than normal reference values); rdw-cv (13.10 + 1.10%) is not different from other stationary alpha - poor, but higher than the normal human group. (4) the Hb of the double heterozygote of hbws compound other static alpha poor is in the normal range (128.60 + 15.67g/l), higher than the hbws composite light beta poor group and light beta - The poor group; MCV (80.27 + 4.70fl) in the critical range, MCH (25.29 + 1.52pg) lower than the normal reference range, but higher than the hbws composite light beta poor group; rdw-cv (13.38 + 1.27%) is lower than the hbws composite light beta poor group, the light beta poor group and the standard type alpha poor. (5) hbws composite light beta poor group HB (112.59 + 14.03g/l), MCV (64.48 + 5.53fl). MCH (19.72 + 1.81pg) was lower than normal range. There was no significant difference between small cell low pigment anemia and light beta poor group Hb, MCV and MCH, rdw-cv (15.67 + 1.34%) was lower than light beta poor group. (6) HB (104.67 + 27.03g/l), MCV (67.76 + 6.10fl), MCH (20.64 + 4.2), - alpha ~ (3.7) group, - alpha ~ (3.7) group. The same expression was small cell low pigmentary anemia, but the Hb level was higher than that of the two groups (P0.05); the MCV in the group of alpha WS alpha /--sea was lower than the alpha CS alpha /--sea group (P0.05), the degree of MCH decreased is the lightest, the rdw-cv increase was the lightest (P0.01). Conclusion: (1) the specificity of the positive diagnosis of the page method region is high, the sensitivity and diagnostic coincidence rate are high and the accuracy is close to the gene. The detection method can be used as a new hbws screening diagnostic method for clinical application. In addition, the page method can also detect the zeta chain for the diagnosis of adult standard alpha poverty. (2) the gene mutation types of hbws in Baise area are mainly alpha WS alpha / alpha alpha, and the hematological phenotype of alpha ws a / ala is basically normal, and it is easy to leak in screening; hbws compound other static alpha. The dual heterozygotes in the poor area were small cells and low pigments, and the hbws composite light beta poor group showed small cell low pigmented anemia; hbh-ws's disease also showed small cell low pigmented anemia, but the degree of anemia was less than - alpha ~ (4.2) /--SEA, - (3.7) /--SEA group light. (3) HbWS different genotypes have their characteristics, but they have their own characteristics. The degree of anemia is lighter than that of the same type of thalassemia. The research results are instructive to the genetic counseling and prenatal diagnosis of the land poverty in this area.
【學(xué)位授予單位】：右江民族醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R556.61
本文編號：2119257
[Abstract]:Objective: (1) to evaluate the accuracy of detection of abnormal hemoglobin Westmead (HbWS) zone by polyacrylamide gel electrophoresis (PAGE), and to provide a new method for screening and diagnosing HbWS from the detection of protein peptide chain. (2) to explore the genotype and hematological phenotypes of HbWS in the population of Baise and the Mediterranean with other different genotypes. Haematological phenotypic difference of anemia (ground poor). Methods: (1) experimental study: in 2015 -2016 years, the native place of Baise region, in our hospital in the poor gene examination, age between 2-60 years of age, gene detection as the gold standard for the diagnosis of HbWS, and the use of PAGE method to detect the HbWS zone with G gamma location with concentration and A gamma position no Strong staining (HbF value) was positive in HbWS area, and the sensitivity and specificity of PAGE positive diagnosis of HbWS were calculated by four grid tables. (2) clinical study: collect the different genotype carriers (patients) from the Baise area in 2010, from the clinic diagnosed as HbWS, and select the Hb according to the inclusion criteria and the exclusion criteria. The carriers (patients) of different types of gene mutations of WS were divided into several control groups (subgroups), and all the samples were carried out by automatic blood cell analyzer, hemoglobin automatic analyzer for hematological analysis, and Gap-PCR detection of deficient alpha and reverse dot print. The RDB method was used to detect the non missing alpha ground poverty and beta poverty. The data were processed with SPSS 19 statistical software. Results: (1) 600 cases were enrolled in the group, 130 cases of HbWS zone positive and 470 cases were diagnosed by PAGE test, 126 cases of HbWS gene mutation were detected by the gold standard method, and 474 cases of.PAG without HbWS gene mutation were found. The diagnostic coincidence rate of E diagnosis HbWS was 98%, sensitivity 96.83%, specificity 98.31%, positive predictive value 0.9385, negative predictive value 0.9915, positive likelihood ratio 57.3, negative likelihood ratio 0.0249. application ROC curve analysis PAGE diagnosis HbWS curve area AUC was 0.942,95% confidence interval (0.893,0.991). (2) collect confirmed HbWS different genotypes group A total of 292 patients (patients) were divided into eight types, including 174 hbws heterozygote (alpha WS alpha / ALA) (59.59%), 43 hbws complex beta poor ground poor (14.73%), 30 double heterozygotes (10.27%), hbh-ws disease 28 (9.59%), and 8 cases (2), hbh-ws disease, and 8 cases (2) (8 cases) with hbws. .74%), 4 cases of hbh-ws disease combined with light beta poor (1.37%), 3 cases of hbws complex beta poor (1.03%) and 2 cases of hbws homozygote (0.68%). (3) the HB (129.68 + 21.11g/l), MCV (86 + 5.36fl), MCH (27.76 + 2.16pg) in the hbws heterozygote group were in the normal range, which was lower than the normal group but higher than the alpha alpha / alpha alpha group, alpha alpha A A / A / alpha / A / - Alpha ~ (3.7) and alpha / - alpha ~ (4.2) groups (the four groups of MCH are lower than normal reference values); rdw-cv (13.10 + 1.10%) is not different from other stationary alpha - poor, but higher than the normal human group. (4) the Hb of the double heterozygote of hbws compound other static alpha poor is in the normal range (128.60 + 15.67g/l), higher than the hbws composite light beta poor group and light beta - The poor group; MCV (80.27 + 4.70fl) in the critical range, MCH (25.29 + 1.52pg) lower than the normal reference range, but higher than the hbws composite light beta poor group; rdw-cv (13.38 + 1.27%) is lower than the hbws composite light beta poor group, the light beta poor group and the standard type alpha poor. (5) hbws composite light beta poor group HB (112.59 + 14.03g/l), MCV (64.48 + 5.53fl). MCH (19.72 + 1.81pg) was lower than normal range. There was no significant difference between small cell low pigment anemia and light beta poor group Hb, MCV and MCH, rdw-cv (15.67 + 1.34%) was lower than light beta poor group. (6) HB (104.67 + 27.03g/l), MCV (67.76 + 6.10fl), MCH (20.64 + 4.2), - alpha ~ (3.7) group, - alpha ~ (3.7) group. The same expression was small cell low pigmentary anemia, but the Hb level was higher than that of the two groups (P0.05); the MCV in the group of alpha WS alpha /--sea was lower than the alpha CS alpha /--sea group (P0.05), the degree of MCH decreased is the lightest, the rdw-cv increase was the lightest (P0.01). Conclusion: (1) the specificity of the positive diagnosis of the page method region is high, the sensitivity and diagnostic coincidence rate are high and the accuracy is close to the gene. The detection method can be used as a new hbws screening diagnostic method for clinical application. In addition, the page method can also detect the zeta chain for the diagnosis of adult standard alpha poverty. (2) the gene mutation types of hbws in Baise area are mainly alpha WS alpha / alpha alpha, and the hematological phenotype of alpha ws a / ala is basically normal, and it is easy to leak in screening; hbws compound other static alpha. The dual heterozygotes in the poor area were small cells and low pigments, and the hbws composite light beta poor group showed small cell low pigmented anemia; hbh-ws's disease also showed small cell low pigmented anemia, but the degree of anemia was less than - alpha ~ (4.2) /--SEA, - (3.7) /--SEA group light. (3) HbWS different genotypes have their characteristics, but they have their own characteristics. The degree of anemia is lighter than that of the same type of thalassemia. The research results are instructive to the genetic counseling and prenatal diagnosis of the land poverty in this area.
【學(xué)位授予單位】：右江民族醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R556.61
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