| Isolation of Fetal Cell from Maternal Blood by Glycophorin A[GPA] Staining and Microdissection. |
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J S Moon, J H Jeong, J M Oh, J D Hwang, S H Yang, J R Roh, S Y Song, S W Kim, J H Lee |
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| Abstract |
OBJECTIVE
Fetal cells in maternal circulation offer a potential source of fetal informations by noninvasive procedure. As there is no completely selective method to separate fetal cells from maternal blood so far, we investigated the effectiveness of isolating nucleated red blood cell[nRBC] from maternal blood by glycophorin A[GPA] staining and microdissection. METHODS: Peripheral blood samples were obtained from 13 pregnant women at 15th to 18th week of gestations. Samples were centrifuged under triple density gradient and handled by magnetic activated cell sorter[MACS] using CD45 monoclonal antibody for removing maternal cells. GPA stain and hematoxylene-eosin counter stain were perfomed to decrease maternal cell contamination. The microdissection of GPA-stained nRBC and nested PCR using Y chromosome-specific primer were underwent for determination of fetal sex. The results were compared with those of amniocentesis for the assessment of efficacy. RESULTS: Four cases had Y-specific sequence on 198 base pair by nested PCR. And other 9 cases did not show Y-specific band. All of Y-band positive cases and additional one of Y-band negative cases were identified to be male fetus by amniocentesis. This study shows 80%[4/5] of sensitivity, 100%[8/8] of specificity, 100% [4/4] of positive and 88.9%[8/9] of negative predictivity. CONCLUSION: These results suggest that GPA staining followed by microdissection could diminish maternal cell contamination and isolate fetal cells effectively. |
| Keywords:
Fetal cell isolation; GPA staining; Microdissection |
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