An Investigation Of Quantitative Fluorescence Quantitative Polymerase Chain Reaction For Detection Trophoblast And JAR Cell From Peripheral Blood

Background Fetomaternal cell traffic in pathological conditions was first recognised a century ago, when the German pathologist, Schmorl identified trophoblasts in the lung capillaries of women dying of eclampsia. In 1969 Walknowska et al demonstrate that fetal cells enter the maternal circulation in normal pregnancy and proposed that these cells had potential for use in chromosome analysis. Since then, several groups claimed Y- chromatin positive cells in maternal blood of pregnant women carrying male fetuses, it suggested that fetal cells in maternal blood could be isolated for non-invasive prenatal diagnosis. But, there were few technique to isolate fetal cells from maternal blood and no sensitive detection. Methods for analysis of fetal cells have little efficiency, It had been found that there are four types of fetal cells in maternal circulation: fetal nucleated red blood cells, trophoblasts, lymphocytes and granulocytes. Trophoblasts have intimate relationship with the utcnis. it is widely accepted that they are present in relatively high numbers in the maternal circulation when they penetrate the uterine mucosa. Trophoblasts can be definitively identified because their unique 5 2001 ~ morphology, secret human chorionic gonadotropin (hCG) and express hCG-mRNA as marker. They are likely ideal fetal cells in maternal circulation for genetic analysis. There are about 10-1000 trophoblasts in maternal blood normally. Factors likely to influence the number of fetal cells circulating include pregnancy-induced hypertension, fetal aneuploidy and.multiple gestation et al. The low fetal cell numbers necessitate removal of maternal cells-enrichment and sensitive methods for quantitative detection fetal cells from the maternal blood. With the advance of molecular technology, Detection non-blood cells or tumor cells in periperal blood had been greatly developed. Some researchers take for mRNA expression as index of diagnosis and surveillance. It is one of the hot spots at present to isolate fetal cells in maternal blood and obtain sufficient genetic information for non-invasive prenatal diagnosis. However, using for clinical diagnosis still has many difficulties. Objective To found sensitive and reliable method for identify fetal trophoblasts or JAR cells from pcriperal blood. Establish an experimental method for non-invasive prenatal diagnosis, investigation physiologic and pathologic significance of trophoblasts existing in maternal blood, the cause of pregnancy-induced hypertension syndrome (PIH) and early diagnosis of metastasis of gestational tumor (Gil?. Method The study was designed to mix known numbers of trophoblasts or JAR cells into lOriil non-pregnant female blood as a model for the rare fetal cells or tumor cells in periperal circulation. After Red Cell Lysis or Ficoll density gradient, total RNA was extracted from the cells, followed by synthesis of cDNA with reverse transcriptase and flurescence quantitative polymerase chain reaction (FQ- 6 2001 ~ PCR)amplification using hCG- P primers and probe. T...