Limits...
Placenta-derived fetal specific mRNA is more readily detectable in maternal plasma than in whole blood.

Heung MM, Jin S, Tsui NB, Ding C, Leung TY, Lau TK, Chiu RW, Lo YM - PLoS ONE (2009)

Bottom Line: The concentrations of placental expressed genes, CSH1, KISS1, PLAC4 and PLAC1 in plasma and whole blood from healthy pregnant and non-pregnant individuals were compared by real-time quantitative reverse-transcriptase polymerase chain reaction analysis.The concentrations of the four transcripts were significantly higher in third trimester maternal whole blood than corresponding plasma without significant correlations.Maternal plasma is preferred over maternal whole blood for placenta-derived fetal RNA detection.

View Article: PubMed Central - PubMed

Affiliation: Centre for Research into Circulating Fetal Nucleic Acids, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong Special Administrative Region, China.

ABSTRACT

Background: Placental mRNA was detected in maternal whole blood, raising the possibility of using maternal blood for noninvasive prenatal diagnosis. We investigated fetal mRNA detection in maternal whole blood and determined if it offered advantages over maternal plasma analysis.

Methodology: The concentrations of placental expressed genes, CSH1, KISS1, PLAC4 and PLAC1 in plasma and whole blood from healthy pregnant and non-pregnant individuals were compared by real-time quantitative reverse-transcriptase polymerase chain reaction analysis. Their fetal specificity was investigated by comparing the transcript concentrations in pre- and post-delivery samples and through SNP genotyping by matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry. The gene expression profiles of pregnant and non-pregnant whole blood were investigated by microarray analysis. Upregulated genes in pregnant whole blood were selected for further quantitative analysis.

Principal findings: The concentrations of the four transcripts were significantly higher in third trimester maternal whole blood than corresponding plasma without significant correlations. KISS1, PLAC4 and PLAC1 were detected in non-pregnant whole blood but not plasma. The transcripts remained detectable in some postpartum whole blood samples. The PLAC4 mRNA in maternal plasma showed fetal genotype while that in corresponding whole blood indicated both fetal and maternal contributions. Microarray analysis revealed upregulation of genes involved in neutrophil functions in pregnant whole blood including DEFA4, CEACAM8, OLFM4, ORM1, MMP8 and MPO. Though possibly pregnancy-related, they were not pregnancy-specific as suggested by the lack of post-delivery reduction in concentrations.

Conclusions: Maternal plasma is preferred over maternal whole blood for placenta-derived fetal RNA detection. Most studied 'placental' mRNA molecules in maternal whole blood were of maternal origin and might be derived from processes such as 'illegitimate transcription'.

Show MeSH

Related in: MedlinePlus

Concentrations of identified whole blood mRNA in third trimester pregnant and non-pregnant whole blood.Box plots of (A) DEFA4 (B) CEACAM8 (C) OLFM4 (D) FLCN (E) ORM1 (F) MMP8 (G) MPO mRNA. The line inside each box denotes the median. The lower and upper limits denote the 25th and 75th percentiles, respectively. The lower and upper whiskers denote the 10th and 90th percentiles, respectively. Filled circles denote the outliers. Asterisks indicate the groups with significant differences in mRNA concentrations of the marker (P<0.05, Kruskal-Wallis test followed by pairwise comparison using the Student-Neuman-Keuls test).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2690655&req=5

pone-0005858-g004: Concentrations of identified whole blood mRNA in third trimester pregnant and non-pregnant whole blood.Box plots of (A) DEFA4 (B) CEACAM8 (C) OLFM4 (D) FLCN (E) ORM1 (F) MMP8 (G) MPO mRNA. The line inside each box denotes the median. The lower and upper limits denote the 25th and 75th percentiles, respectively. The lower and upper whiskers denote the 10th and 90th percentiles, respectively. Filled circles denote the outliers. Asterisks indicate the groups with significant differences in mRNA concentrations of the marker (P<0.05, Kruskal-Wallis test followed by pairwise comparison using the Student-Neuman-Keuls test).

Mentions: When comparing the absolute concentrations of DEFA4, CEACAM8, OLFM4, FLCN, ORM1, MMP8 and MPO mRNA in whole blood samples from third trimester pregnant, non-pregnant female and male, statistical significant differences were observed in the markers (P<0.001, Kruskal-Wallis ANOVA test) except FLCN (Pā€Š=ā€Š0.231, Kruskal-Wallis ANOVA test) (Figure 4).


Placenta-derived fetal specific mRNA is more readily detectable in maternal plasma than in whole blood.

Heung MM, Jin S, Tsui NB, Ding C, Leung TY, Lau TK, Chiu RW, Lo YM - PLoS ONE (2009)

Concentrations of identified whole blood mRNA in third trimester pregnant and non-pregnant whole blood.Box plots of (A) DEFA4 (B) CEACAM8 (C) OLFM4 (D) FLCN (E) ORM1 (F) MMP8 (G) MPO mRNA. The line inside each box denotes the median. The lower and upper limits denote the 25th and 75th percentiles, respectively. The lower and upper whiskers denote the 10th and 90th percentiles, respectively. Filled circles denote the outliers. Asterisks indicate the groups with significant differences in mRNA concentrations of the marker (P<0.05, Kruskal-Wallis test followed by pairwise comparison using the Student-Neuman-Keuls test).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2690655&req=5

pone-0005858-g004: Concentrations of identified whole blood mRNA in third trimester pregnant and non-pregnant whole blood.Box plots of (A) DEFA4 (B) CEACAM8 (C) OLFM4 (D) FLCN (E) ORM1 (F) MMP8 (G) MPO mRNA. The line inside each box denotes the median. The lower and upper limits denote the 25th and 75th percentiles, respectively. The lower and upper whiskers denote the 10th and 90th percentiles, respectively. Filled circles denote the outliers. Asterisks indicate the groups with significant differences in mRNA concentrations of the marker (P<0.05, Kruskal-Wallis test followed by pairwise comparison using the Student-Neuman-Keuls test).
Mentions: When comparing the absolute concentrations of DEFA4, CEACAM8, OLFM4, FLCN, ORM1, MMP8 and MPO mRNA in whole blood samples from third trimester pregnant, non-pregnant female and male, statistical significant differences were observed in the markers (P<0.001, Kruskal-Wallis ANOVA test) except FLCN (Pā€Š=ā€Š0.231, Kruskal-Wallis ANOVA test) (Figure 4).

Bottom Line: The concentrations of placental expressed genes, CSH1, KISS1, PLAC4 and PLAC1 in plasma and whole blood from healthy pregnant and non-pregnant individuals were compared by real-time quantitative reverse-transcriptase polymerase chain reaction analysis.The concentrations of the four transcripts were significantly higher in third trimester maternal whole blood than corresponding plasma without significant correlations.Maternal plasma is preferred over maternal whole blood for placenta-derived fetal RNA detection.

View Article: PubMed Central - PubMed

Affiliation: Centre for Research into Circulating Fetal Nucleic Acids, Li Ka Shing Institute of Health Sciences, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong Special Administrative Region, China.

ABSTRACT

Background: Placental mRNA was detected in maternal whole blood, raising the possibility of using maternal blood for noninvasive prenatal diagnosis. We investigated fetal mRNA detection in maternal whole blood and determined if it offered advantages over maternal plasma analysis.

Methodology: The concentrations of placental expressed genes, CSH1, KISS1, PLAC4 and PLAC1 in plasma and whole blood from healthy pregnant and non-pregnant individuals were compared by real-time quantitative reverse-transcriptase polymerase chain reaction analysis. Their fetal specificity was investigated by comparing the transcript concentrations in pre- and post-delivery samples and through SNP genotyping by matrix-assisted laser-desorption and ionization time-of-flight mass spectrometry. The gene expression profiles of pregnant and non-pregnant whole blood were investigated by microarray analysis. Upregulated genes in pregnant whole blood were selected for further quantitative analysis.

Principal findings: The concentrations of the four transcripts were significantly higher in third trimester maternal whole blood than corresponding plasma without significant correlations. KISS1, PLAC4 and PLAC1 were detected in non-pregnant whole blood but not plasma. The transcripts remained detectable in some postpartum whole blood samples. The PLAC4 mRNA in maternal plasma showed fetal genotype while that in corresponding whole blood indicated both fetal and maternal contributions. Microarray analysis revealed upregulation of genes involved in neutrophil functions in pregnant whole blood including DEFA4, CEACAM8, OLFM4, ORM1, MMP8 and MPO. Though possibly pregnancy-related, they were not pregnancy-specific as suggested by the lack of post-delivery reduction in concentrations.

Conclusions: Maternal plasma is preferred over maternal whole blood for placenta-derived fetal RNA detection. Most studied 'placental' mRNA molecules in maternal whole blood were of maternal origin and might be derived from processes such as 'illegitimate transcription'.

Show MeSH
Related in: MedlinePlus