1991 Fiscal Year Final Research Report Summary
Development of prenatal diagnosis of rubella infection with DNA
| Project/Area Number |
01571274
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | National Institute of Health |
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Principal Investigator |
KATOW Shigetaka National Institute of Health, Department of Measles Virus, Chief Researcher, 麻紳ウイルス部, 主任研究官 (20211162)
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| Co-Investigator(Kenkyū-buntansha) |
TANABAYASHI Kiyoshi National Institute of Health, Department of Measles Virus, Researcher, 麻疹ウイルス部, 研究員 (50197505)
SUZUMORI Kaoru Nagoya City University Medical School, Department of Obstetrics & Gynecology, As, 医学部, 助教授 (80117829)
KAWANA Takashi Tokyo University Branch Hospital, Department of Obstetrics & Gynecology, Profess, 医学部付属病院・分院・産婦人科, 教授 (90010272)
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| Project Period (FY) |
1989 – 1991
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| Keywords | Rubella Virus / Congenital Rubella Syndrome (CRS) / Polymerase Chain Reaction (PCR) / Pregnancy / Prenatal Diagnosis / Virus Genome / Genome Diagnosis / Chorionic Villi Sampling (CVS) |
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| Research Abstract |
Primary infection with rubella virus during early pregnancy often leads to intrauterine malformations. It is very important to establish the evidence of virus infection in the early gestation by confirmatory laboratory diagnosis. As a result of the ambiguity of serological diagnosis, artificial abortion was adopted in many cases for fear of having babies with congenital malformation. Definite diagnosis of the intrauterine infection, therefore, has been required to avoid unnecessary artificial termination.
We developed simple method for detection of rubelia virus genome in a variety of maternal and fetal tissues. Total tissue RNA extracted by an acid guanidium-thiocyanate-phenol-chloroform method was reverse transcribed in the presence of a set of synthetic primers then amplified through PCR using the same primers. The resulted DNA was subjected to the second PCR using a different set of primers. The PCR products were then electrophoresed in an agarose gel followed by staining with ethidium bromide.
DNA fragments of expected size were amplified from tissues of fetal origin, such as, chorionic villi, amniotic fluids, and umbilical cord blood lymphocytes obtained by biopsy from pregnant women infected with rubella. The PCR products were proven to be specific to rubella virus by direct sequencing of the products.
By the development of reliable diagnostic method of virus infection in fetus based on viral genome detection in addition to serological methods, we could expect reduction in numbers of babies with congenital rubella syndrome and of unnecessary artificial abortion of healthy babies.
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