应用多色探针熔解曲线法检测G6PD缺乏症杂合子的基因突变

胡韦维; 李进; 刘益; 刘之岱; 张娟; 余朝文; 邹琳; 张鹏辉

doi:10.16016/j.1000-5404.201710014

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应用多色探针熔解曲线法检测G6PD缺乏症杂合子的基因突变

临床医学 | 更新时间：2026-01-09

- 应用多色探针熔解曲线法检测G6PD缺乏症杂合子的基因突变
- Detection of gene mutation in heterozygous of glucose-6-phosphate dehydrogenase deficiency by multicolor melting curve analysis
- 第三军医大学学报 2018年40卷第9期页码：830-835
- 作者机构：
  
  1.400014 重庆, 重庆医科大学附属儿童医院检验科, 儿童发育疾病研究教育部重点实验室, 儿童发育重大疾病国家国际科技合作基地, 重庆市干细胞治疗工程技术研究中心
  2.400042 重庆, 陆军军医大学(第三军医大学)第三附属医院(野战外科研究所)检验科
  3.400014 重庆, 重庆医科大学附属儿童医院临床分子医学中心
- 作者简介：
  
  [张鹏辉, 电话:(023)63631417, E-mail:zhangph7203@sina.com ZHANG Penghui, Tel:86-23-63631417, E-mail:zhangph7203@sina.com
- 基金信息：
  
  重庆市社会民生科技创新专项(CSTC2016shmszx130032)
- DOI：10.16016/j.1000-5404.201710014
  中图分类号： R394.2;R446.9;R722.11
- 收稿：2017-10-09，
  
  修回：2018-02-02，
  
  纸质出版：2018-05-15
- 稿件说明：
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胡韦维, 李进, 刘益, 等. 应用多色探针熔解曲线法检测G6PD缺乏症杂合子的基因突变[J]. 第三军医大学学报, 2018,40(9):830-835.

Weiwei HU, Jin LI, Yi LIU, et al. Detection of gene mutation in heterozygous of glucose-6-phosphate dehydrogenase deficiency by multicolor melting curve analysis[J]. Journal of Third Military Medical University, 2018, 40(9): 830-835.
胡韦维, 李进, 刘益, 等. 应用多色探针熔解曲线法检测G6PD缺乏症杂合子的基因突变[J]. 第三军医大学学报, 2018,40(9):830-835. DOI： 10.16016/j.1000-5404.201710014.

Weiwei HU, Jin LI, Yi LIU, et al. Detection of gene mutation in heterozygous of glucose-6-phosphate dehydrogenase deficiency by multicolor melting curve analysis[J]. Journal of Third Military Medical University, 2018, 40(9): 830-835. DOI： 10.16016/j.1000-5404.201710014.

摘要

目的

应用多色探针熔解曲线法(multicolor melting curve analysis

MMCA)

建立一种快速筛查葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症杂合子的临床方法。

方法

收集重庆地区2015年1月至2016年7月共429例(男性342例

女性87例)疑似G6PD缺乏症的患者外周血

采用G6PD/6PGD定量比值法和MMCA法分别检测男性和女性G6PD酶活性和基因突变

以金标准Sanger测序来评价两种方法。同时收集重庆地区进行G6PD缺乏症新生儿疾病筛查的1 754份女性末梢血滤纸片

采用MMCA法筛查G6PD基因突变。

结果

429例疑似病例中

G6PD/6PGD定量比值法诊断男性的灵敏度96.8%、特异性100%

与Sanger测序法一致性好(

Kappa

=0.917

0.05);但女性的灵敏度仅10%

特异性100%

与Sanger测序法一致性差(

Kappa

=0.127

0.05)。MMCA法诊断男性的灵敏度95.7%

特异性100%

与Sanger测序法一致性较好(

Kappa

=0.891

0.05);女性的灵敏度100%

特异性100%

与Sanger测序法结果完全吻合(

Kappa

=1.000

0.05)。1 754例女性中杂合子占1.14%(20/1 754)

G6PD酶活性均正常。

结论

与传统的G6PD/6PGD定量比值法比较

MMCA法检测杂合子灵敏度更高

可用于G6PD缺乏症杂合子筛查

为一种简便、准确的诊断新方法。

Abstract

Objective

To establish a rapid clinical method for detecting the heterozygote with glucose-6-phosphate dehydrogenase (G6PD) deficiency by multicolor melting curve analysis (MMCA).

Methods

A total of 429 children (342 males and 87 females) suspected with G6PD deficiency who lived in Chongqing region and visited our Children's Hospital between January 2015 and July 2016 were collected in this study.G6PD/6PGD quantitative ratio test and MMCA were applied respectively to determine the enzyme activity and G6PD gene mutation

and Sanger sequencing was also employed as gold standard method.At the same time

totally 1 754 peripheral blood filter papers from women recruited from Chongqing region were identified by MMCA for G6PD deficiency disease screening.

Results

Among the 429 cases suspected with G6PD deficiency

G6PD/6PGD ratio test showed the sensitivity and specificity were 96.8% and 100% respectively for the male

which were consistent with the results of Sanger sequencing (

Kappa

=0.917

0.05);while the sensitivity and specificity were only 10% and 100% for the female

inconsistent with the results of Sanger sequencing (

Kappa

=0.127

0.05).MMCA showed the sensitivity and specificity were 95.7% and 100% respectively for the male

consistent with Sanger sequencing (

Kappa

=0.891

0.05);and were 100% and 100% for the female

which were in good agreement with Sanger sequencing (

Kappa

=1.000

0.05).The heterozygotes accounted for 1.14% (20/1 754)

with normal activity of G6PD enzyme.

Conclusion

Compared with the traditional method quantitative ratio of G6PD/6PGD

MMCA assay can screen the heterozygote with higher sensitivity

and is a simple and accurate new diagnostic method.

关键词

Keywords

references

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