Quantitative polymerase chain reaction assessment of chimerism in non-human primates after sex-mismatched islet and bone marrow transplantation.
Quantitative polymerase chain reaction assessment of chimerism in non-human primates after sex-mismatched islet and bone marrow transplantation.
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性别不匹配的胰岛和骨髓移植后非人灵长类动物嵌合的定量聚合酶链反应评估。
DOI：
10.1097/00007890-200004270-00031
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发表时间：
2000
期刊：
影响因子：
6.2
通讯作者：
Kenyon,NS
中科院分区：
文献类型：
--
作者：
Han,D;Ricordi,C;Xu,X;Kenyon,NS
背景：准确评估胰岛和骨髓移植(BMT)受者的嵌合体可能有助于更清楚地评估嵌合体在胰岛植入或排斥反应中的作用。建立了一种定量聚合酶链式反应(PCR)技术，用于检测同供者的异基因男性胰岛和椎体骨髓(VBM)中Y染色体性别决定区域(SRY)的性别决定区(SRY)。每个DNA样本都与恒定量的IS进行共扩增。通过绘制SRY和IS-PCR产物的强度比与已知男性DNA输入百分比的对数的回归方程，计算出测试样本中男性DNA的浓度。结果该方法修正了PCR技术效率的变异性，也克服了竞争PCR耗时的缺点。利用这一方法，我们对雄性胰岛和VBM的雌性狒狒受体的样本中雄性DNA的数量进行了定量。在骨髓移植后一天采集的样本中有可检测到的男性供体DNA；骨髓移植前样本为阴性。该技术同样适用于恒河猴和食蟹猴的样本。结论：这是一种在非人类灵长类动物模型中准确评估性别不匹配异基因骨髓移植后嵌合体的实用方法。
Background.Accurate assessment of chimerism in recipients of islet and bone marrow transplantation (BMT) may allow for a clearer assessment of the role of chimerism in islet engraftment or rejection. A quantitative polymerase chain reaction (PCR) assay was developed for the detection of the sex-determining region of the Y chromosome (SRY) in peripheral blood samples from female non-human primate recipients of allogeneic male islets and vertebral body marrow (VBM) from the same donor.Methods.The assay incorporates a synthetic internal standard (IS) containing the same primer template sequences as the target to compete for primer annealing and amplification. Each DNA sample was coamplified with a constant amount of IS. The concentration of male DNA in the test samples was calculated from the regression equation of a standard curve that was generated by plotting the logarithm of the ratio of the intensities of SRY to IS PCR products versus the logarithm of known percentages of input male DNA.Results.This method allows for a correction of the variability of efficiency of the PCR technique and also overcomes the drawback of time-consuming competitive PCR. Using this assay, we quantitated the amount of male DNA in samples taken from female baboon recipients of male islets and VBM. There was detectable male donor DNA in the samples taken one day after BMT; pre-BMT samples were negative. This technique works well for samples obtained from rhesus and cynomogus monkeys as well.Conclusions.It is a practical method for accurately evaluation of chimerism after sex-mismatched allogeneic BMT in non-human primate models.
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DOI：
10.1101/gr.3.6.361
发表时间：
1994
期刊：
PCR methods and applications
影响因子：
--
作者：
Sylvie Patri;Laurence Oaheron;Alain Kitzis;J.
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DOI：
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发表时间：
1993
期刊：
Cytogenetics and cell genetics
影响因子：
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作者：
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影响因子：
14.9
作者：
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影响因子：
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影响因子：
6.2
作者：
Pavlakis, M;Strehlau, J;Strom, TB
通讯作者：
Strom, TB