PMID- 12533814
OWN - NLM
STAT- MEDLINE
DCOM- 20030702
LR  - 20061115
IS  - 0197-3851 (Print)
IS  - 0197-3851 (Linking)
VI  - 23
IP  - 1
DP  - 2003 Jan
TI  - Detection of gamma-globin mRNA in fetal nucleated red blood cells by PNA 
      fluorescence in situ hybridization.
PG  - 52-9
AB  - OBJECTIVES: Fetal nucleated red blood cells (NRBC) that enter the peripheral 
      blood of the mother are suitable for non-invasive prenatal diagnosis. The 
      application of peptide nucleic acid (PNA) probes for tyramide amplified flow 
      fluorescence in situ hybridization (FISH) detection of gamma-globin mRNA in fixed 
      fetal NRBC is investigated. METHODS: Hemin-induced K562 cells or nucleated blood 
      cells (NBC) from male cord blood were mixed with NBC from non-pregnant women and 
      analysed using both slide and flow FISH protocols. Post-chorionic villus sampling 
      (CVS) blood samples from pregnant females carrying male fetuses were flow-sorted 
      (2 x 10(6) NBC/sample). Y chromosome-specific PNA FISH was used to confirm that 
      the identified gamma-globin mRNA stained cells were of fetal origin. RESULTS: 
      Flow FISH isolated gamma-globin mRNA positive NBCs showing characteristic 
      cytoplasmic staining were all Y positive. The amplification system generated a 
      population of false positive cells that were, however, easy to distinguish from 
      the NRBCs in the microscope. CONCLUSION: The gamma-globin mRNA specific PNA 
      probes can be used for detection and isolation of fetal NRBCs from maternal 
      blood. The method has additional potential for the study of gamma-globin mRNA 
      levels or the frequency of adult NRBC (F cells) in patients with 
      hemoglobinopathies.
CI  - Copyright 2003 John Wiley & Sons, Ltd.
FAU - Larsen, Rasmus Dines
AU  - Larsen RD
AD  - DakoCytomation A/S, Produktionsvej 42, DK-2600 Glostrup, Copenhagen, Denmark. 
      rasmuslarsen@mail.dk
FAU - Schonau, Andreas
AU  - Schonau A
FAU - Thisted, Marianne
AU  - Thisted M
FAU - Petersen, Kenneth Heesche
AU  - Petersen KH
FAU - Lohse, Jesper
AU  - Lohse J
FAU - Christensen, Britta
AU  - Christensen B
FAU - Philip, John
AU  - Philip J
FAU - Pluzek, Karl-Johan
AU  - Pluzek KJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Prenat Diagn
JT  - Prenatal diagnosis
JID - 8106540
RN  - 0 (Immunoglobulin G)
RN  - 0 (Peptide Nucleic Acids)
RN  - 0 (RNA, Messenger)
SB  - IM
MH  - Adult
MH  - Base Sequence
MH  - Chromosomes, Human, Y
MH  - Erythroblasts/*chemistry/cytology
MH  - Female
MH  - Fetal Blood/*chemistry/cytology
MH  - Fetomaternal Transfusion
MH  - Flow Cytometry
MH  - Humans
MH  - Immunoglobulin G/genetics
MH  - *In Situ Hybridization, Fluorescence
MH  - Infant, Newborn
MH  - Male
MH  - Microscopy, Fluorescence
MH  - Molecular Sequence Data
MH  - Peptide Nucleic Acids/*analysis
MH  - Pregnancy
MH  - *Prenatal Diagnosis/*methods
MH  - RNA, Messenger/*analysis
MH  - Sequence Alignment
MH  - Tumor Cells, Cultured
EDAT- 2003/01/21 04:00
MHDA- 2003/07/03 05:00
CRDT- 2003/01/21 04:00
PHST- 2003/01/21 04:00 [pubmed]
PHST- 2003/07/03 05:00 [medline]
PHST- 2003/01/21 04:00 [entrez]
AID - 10.1002/pd.520 [doi]
PST - ppublish
SO  - Prenat Diagn. 2003 Jan;23(1):52-9. doi: 10.1002/pd.520.