Diagnosis of heterozygous states for adenine phosphoribosyltransferase deficiency based on detection of in vivo somatic mutants in blood T cells: application to screening of heterozygotes.
Diagnosis of heterozygous states for adenine phosphoribosyltransferase deficiency based on detection of in vivo somatic mutants in blood T cells: application to screening of heterozygotes.
复制标题
基于血液 T 细胞体内体细胞突变体检测的腺嘌呤磷酸核糖基转移酶缺陷杂合状态诊断：应用于杂合子筛选。
DOI：
--
复制
发表时间：
1991
影响因子：
9.8
通讯作者：
N. Kamatani
中科院分区：
文献类型：
--
作者：
M. Hakoda;Hisashi Yamanaka;N. Kamatani;N. Kamatani
准确诊断具有未知突变的常染色体隐性遗传病杂合子可能很困难。利用体内发生的独特现象，我们设计了一种诊断杂合子腺嘌呤磷酸核糖转移酶（APRT）缺陷的方法，该方法为正常和杂合子受试者之间的定性区分铺平了道路。我们用 2,6-二氨基嘌呤（一种 APRT 依赖性细胞毒素）培养外周血单核细胞，以寻找体内突变细胞。经检查，发现 15 个假定的杂合子以相当高的频率拥有此类突变细胞；因此，假阴性诊断的可能性不大。对来自两个杂合子的 82 个抗性克隆的基因组 DNA 分析表明，64 个（78%）已经失去了萌发时完整的等位基因。对 APRT 缺陷家庭的 13 名成员进行了检查；八个被证明是杂合子。在来自日本两个不同住宅区的 425 名个体中，发现了两个杂合子。对于两个杂合子，通过两种不同的方法验证了杂合性的真实性；因此，可以排除假阳性诊断。我们的数据显示计算出的杂合子频率为 0.47%（95% 置信限；0.05%-1.7%），该值与根据 2,8-二羟基腺嘌呤尿石症发病率数据计算得出的值 (1.2%) 一致。这种用于识别杂合子的新颖遗传方法目前正在测试中，以寻找人类中其他酶的缺陷。
An accurate diagnosis of heterozygotes for autosomal recessive disorders with unknown mutations can be difficult. Using a unique phenomenon occurring in vivo, we designed a method for the diagnosis of heterozygotes for adenine phosphoribosyltransferase (APRT) deficiency which makes way for a qualitative distinction between normal and heterozygous subjects. We cultured peripheral blood mononuclear cells with 2,6-diaminopurine, an APRT-dependent cytotoxin, to search for in vivo mutational cells. Fifteen putative heterozygotes examined were found to possess such mutant cells at rather high frequencies; thus, a false negative diagnosis is unlikely. The analysis of genomic DNA in 82 resistant clones from two of the heterozygotes clarified that 64 (78%) had lost the germinally intact alleles. Thirteen members of APRT-deficient families were examined; eight proved to be heterozygotes. Among 425 individuals from two separate residential areas of Japan, two heterozygotes were found. The authenticity of the heterozygosity was validated by two separate methods for the two heterozygotes; hence, a false positive diagnosis can be ruled out. Our data showed a calculated heterozygote frequency of 0.47% (95% confidence limits; 0.05%-1.7%), a value compatible with that (1.2%) calculated from data concerning the incidence of 2,8-dihydroxyadenine urolithiasis. This novel genetic approach for identifying heterozygotes is now being tested to search for other enzyme deficiencies in humans.
DOI：
10.1016/s0021-9258(19)62682-7
发表时间：
1986-05
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
B. Hove-Jensen;K. Harlow;C. King;R. Switzer
通讯作者：
B. Hove-Jensen;K. Harlow;C. King;R. Switzer
人 T 淋巴细胞体内 hprt 突变的分子分析。
影响因子：
2.7
作者：
Nicklas,JA;Hunter,TC;Sullivan,LM;Berman,JK;O'Neill,JP;Albertini,RJ
通讯作者：
Albertini,RJ
人 T 淋巴细胞体内 hprt 突变的分子分析。
影响因子：
2.7
作者：
Nicklas,JA;Falta,MT;Hunter,TC;O'Neill,JP;Jacobson-Kram,D;Williams,JR;Albertini,RJ
通讯作者：
Albertini,RJ