Reagent for multiplex real-time quantitative method for evaluating hemopoietic stem cell transplantation state
A hematopoietic stem cell, real-time quantitative technology, applied in the direction of microbial measurement/testing, biochemical equipment and methods, etc., can solve the problems of non-disclosure and inapplicability to the Chinese population
Active Publication Date: 2017-08-04
浙江省血液中心
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Problems solved by technology
At present, foreign countries have developed polynucleotide polymorphism quantitative methods for the Caucasian population, but due to differences in race and genetics, these nucleotide sites are not suitable for the Chinese population, and the primers and probes for these sites Needles and methods are protected by foreign patents and are not disclosed
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[0118] The content of the present invention will be described in further detail below in conjunction with the examples.
[0119] This implementation specifically takes the analysis of cell chimerism in patients after hematopoietic stem cell transplantation as an example to describe the content of the present invention in detail. The specific steps of a multiple real-time quantitative method for assessing the implantation status of hematopoietic stem cell transplantation adopted by the present invention are as follows:
[0120] 1. DNA template preparation
[0121] Genomic DNA from three blood samples of the patient before transplantation, the donor, and the patient after transplantation were prepared as templates for PCR amplification in subsequent steps. Take 200 μl each of the three whole blood samples to be tested, extract genomic DNA according to the instructions of the QuickGene DNA whole blood kit kit, and measure the DNA concentration and purity with a spectrophotometer...
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The invention discloses a reagent for a multiplex real-time quantitative method for evaluating a hemopoietic stem cell transplantation state. The reagent contains 26 pairs of primers of an amplified 673 site insertion allele, a 673 site deletion allele, a 688 site insertion allele, a 688 site deletion allele, an 803 site insertion allele, an 803 site deletion allele, a 918 site insertion allele, a 918 site deletion allele, a 944 site insertion allele, a 944 site deletion allele, a 1066 site insertion allele, a 1066 site deletion allele, a 1100 site insertion allele, a 1100 site deletion allele, a 1151 site insertion allele, a 1151 site deletion allele, a 1625 site insertion allele, a 1625 site deletion allele, a 1989 site insertion allele, a 1989 site deletion allele, a 2504 site insertion allele, a 2504 site deletion allele, a 2568 site insertion allele, a 2568 site deletion allele, a 2676 site insertion allele and a 2676 site deletion allele, and 13 oligonucleotide probes for fluorescence detection. The reagent is of great significance to improvement of the hemopoietic stem cell transplantation survival rate and the postoperative survival rate.
Description
technical field [0001] The invention relates to a reagent used in a gene quantitative detection method, in particular to a reagent used in a multiple real-time quantitative method for evaluating the engraftment state of hematopoietic stem cell transplantation. Background technique [0002] Allogeneic hematopoietic stem cell transplantation (Hematopoiet ic stem cell transplantation, HSCT), including bone marrow transplantation, umbilical cord blood stem cell transplantation, peripheral blood stem cell transplantation, etc., is the treatment of malignant blood diseases (such as leukemia) and severe non-malignant blood diseases (such as severe An important means for aplastic anemia, severe combined immunodeficiency, etc.). However, all forms of transplantation have the possibility of disease recurrence. Taking leukemia as an example, even with high-intensity "myeloablative" pretreatment, it is difficult to completely eliminate the patient's own leukemia cell clones. Malignant...
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IPC IPC(8): C12Q1/68
CPCC12Q1/6851C12Q2531/113C12Q2563/107
Inventor 许先国何吉洪小珍章伟朱发明
Owner 浙江省血液中心