| Project/Area Number |
10671027
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Embryonic/Neonatal medicine
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| Research Institution | Kanazawa Medical University |
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Principal Investigator |
TAKABAYASHI haruo Kanazawa Medical University Medical Research Institute, Associate Professor, 総合医学研究所, 助教授 (60171542)
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| Co-Investigator(Kenkyū-buntansha) |
FUJII Ryota Kanazawa Medical University, Dept.of Ob/Gyn, Assistant, 医学部, 助手 (30298351)
IURA Toshihiko Kanazawa Medical University, Dept.of Ob/Gyn, Lecturer, 医学部, 講師 (40202840)
TANINO Mikio Kanazawa Medical University, Dept.of Pathology, Associate Professor, 医学部, 助教授 (60135051)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1999: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1998: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | maternal blood / nucleated erythrocyte / fetal diagnosis / DNA diagnosis / micromanipulation / PEP / TaqMan PCR / FISH / マイクロマニビュレーション / マイクロマニピュレーション |
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| Research Abstract |
The development of a real-time quantitative TaqMan PCR assay permits the high quality analysis of DNA or RNA. With the TaqMan PCR, Lo.et al. (1998) were successful in measuring the concentration of fetal DNA in maternal plasma and serum.
Using ABI PRISM 7700 System (PE Applied Biosystems), we tried to detect the target DNA sequence of SRY and beta actin locus in the PEP (primer extension preamplification) products from a single cell. Single NRBCs (nucleated red blood cells) were retrieved from cord blood smear of male fetus using a micromanipulator.
The detection rates of beta actin and SRY locus were 55% and 35%, respectively. Quantitative results of PEP products from single NRBCs were 0.5-59.5 (m=4.8) copies in beta actin and 2.4-278.4 (m=17.8) copies in SRY locus.
Our quantitative data show the concentration of a single locus DNA amplified by PEP from a single NRBC. The advantages of PEP-TaqMan PCR system are high specificity and sensitivity.
This system seems a powerful tool for fetal DNA diagnosis from maternal blood.
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