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Publication
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| Research Title |
Prenatal diagnosis of severe thalassemia diseases using direct PCR on amniotic fluid specimen |
| Date of Distribution |
11 May 2018 |
| Conference |
| Title of the Conference |
ISLH 2018 |
| Organiser |
the International Society for Laboratory Hematology (ISLH) and the Belgian Society on Thrombosis and Haemostasis (BSTH) |
| Conference Place |
SQUARE Meeting Centre, Brussels, Belgium |
| Province/State |
Brussels, Belgium |
| Conference Date |
9 May 2018 |
| To |
12 May 2018 |
| Proceeding Paper |
| Volume |
- |
| Issue |
- |
| Page |
- |
| Editors/edition/publisher |
- |
| Abstract |
Introduction: Thalassemias are common in Thailand and other Southeast Asiancountries. Severe forms of thalassemia targeted at a prevention and control program include homozygous α0-thalassemia, homozygous β-thalassemia and β0-thalassemia/Hb E. Prenatal diagnosis of the at-risk couple is an important step for preventing these diseases. Currently, this is done mostly by amniocentesis and DNA analysis. In order to simplify this process of prenatal diagnosis, we have developed a direct PCR protocol on amniotic fluid specimen.
Methods: One mL of amniotic fluid specimen is centrifuged at 3500 rpm for 10 min and supernatant fluid was discarded. Three µL of amniotic fluid pellets was used for direct PCR in a high pH PCR buffer developed.The direct PCR protocol was tested for identifying α0-thalassemia, β0-thalassemia and Hb E in 144 amniotic fluid specimens in blinded trial and results compared with conventional method on purified DNA specimen.
Results: Among 144 specimens,94 fetuses were at-risk of having homozygous α0-thalassemia. Analysis using direct PCR identified 8 affected fetuses, 50 α0-thalassemia carriers and 36 unaffected fetuses. Among 49 fetuses at-risk for β0-thalassemia/Hb E and 1 fetus at-risk for both β0-thalassemia/Hb E and homozygous β0-thalassemia, direct PCR identified 10 affected fetuses (including 9 β0-thalassemia/Hb E and 1 homozygous β0-thalassemia), 11 thalassemia carriers and 29 unaffected fetuses. A 100% concordance results with conventional PCR on purified DNA specimens were observed.
Conclusions: The developed protocol for direct PCR on amniotic fluid specimen is simpler, rapid and reliable. The method requires small amounts of uncultured amniotic fluid specimen and could be applied effectively to prenatal diagnosis of the 3 severe thalassemia diseases. This should prove useful in a prevention and control program of thalassemia in the region. |
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| Peer Review Status |
มีผู้ประเมินอิสระ |
| Level of Conference |
นานาชาติ |
| Type of Proceeding |
Abstract |
| Type of Presentation |
Poster |
| Part of thesis |
true |
| Presentation awarding |
false |
| Attach file |
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| Citation |
0
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Journal Publication
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