Fluctuation of Maternal and Fetal Free Extracellular Circulatory DNA in Maternal Plasma

Zhong, Xiao Yan; Burk, M.; Troeger, Carolyn; Kang, Anjeung; Holzgreve, Wolfgang; Hahn, Sinuhe

doi:10.1097/00006250-200012000-00023

Cited by 22 publications

(27 citation statements)

References 18 publications

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“…Our results indicate that there are between 2 and 6 fetal nucleated cells per milliliter of maternal blood in pregnant women bearing a normal male fetus during the period from 18 to 22 weeks of pregnancy. Using real‐time quantitative PCR technology, Zhong et al [17] observed a mean of 2.2‐fold within individual fluctuation of free extra‐cellular fetal DNA after repeated sampling of pregnant women. If free extra‐cellular fetal DNA is proportional to the number of circulating fetal cells (assuming similar apoptosis rates for most pregnant women), the 3‐fold variation we observed between women might be due to the intra‐individual day‐to‐day variation.…”

Section: Discussionmentioning

confidence: 99%

Quantification of all fetal nucleated cells in maternal blood between the 18th and 22nd weeks of pregnancy using molecular cytogenetic techniques

et al. 2001

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Different types of nucleated fetal cells (trophoblasts, erythroblasts, lymphocytes, and granulocytes) have been recovered in maternal peripheral blood. In spite of many attempts to estimate the number of fetal cells in maternal circulation, there is still much controversy concerning this aspect. The numbers obtained vary widely, ranging from 1 nucleated cell per 104 to 1 per 109 nucleated maternal cells. The purpose of our project was to determine the absolute number of all different types of male fetal nucleated cells per unit volume of peripheral maternal blood. Peripheral blood samples were obtained from 12 normal pregnant women known to carry a male fetus between 18 and 22 weeks of pregnancy. Three milliliters (3 ml) of maternal blood has been processed without any enrichment procedures. Fluorescence in situ hybridization (FISH) and primed in situ labeling (PRINS) were performed, and fetal XY cells were identified (among maternal XX cells) and scored by fluorescent microscopy screening. The total number of male fetal nucleated cells per milliliter of maternal blood was consistent in each woman studied and varied from 2 to 6 cells per milliliter within the group of normal pregnancies. The number of fetal cells in maternal blood, at a given period, is reproducible and can therefore be assessed by cytogenetic methods. This confirms the possibility of developing a non-invasive prenatal diagnosis test for aneuploidies. Furthermore, we demonstrate that it is possible to repeatedly identify an extremely small number of fetal cells among millions of maternal cells.

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Section: Discussionmentioning

confidence: 99%

Quantification of all fetal nucleated cells in maternal blood between the 18th and 22nd weeks of pregnancy using molecular cytogenetic techniques

et al. 2001

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“…The question is, how many times is it necessary to test before reaching reasonable certainty? Published studies reveal up to eight runs of each sample [21] and, as false‐positive results are encountered as well as false negatives [22,23], certainty may be hard to achieve. For the fetus, the consequence of a false‐negative RHD determination may be more severe than a false‐positive result, as the latter would only result in unnecessary prophylaxis.…”

Section: Discussionmentioning

confidence: 99%

“…A study by Zhong et al . [23] reported a fluctuation of up to 4·5 times in free fetal DNA over a 3‐day period. The explanation for this individual variation is not clear.…”

Section: Discussionmentioning

confidence: 99%

Prenatal genotyping ofRHDandSRYusing maternal blood

et al. 2003

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“…The white cells in whole blood provide high quantities of extractable genomic DNA, primarily from the mother, whereas serum contains cell‐free DNA of generally low concentration (Kaufman, Wu, Donghern, & Cseke, ). In addition, cell‐free DNA concentrations in serum can vary substantially among individuals, and fluctuates when measured over both long and short periods (Stroun et al, ; Zhong et al, ). Among rhinoceroses, cell‐free DNA yield and PCR amplification quality varied by individual and by species; indeed, overall PCR product quality was low or absent from Sumatran rhinoceros compared to the other African and Asian male rhinoceroses examined.…”

Section: Discussionmentioning

confidence: 99%

Early fetal sexing in the rhinoceros by detection of male‐specific genes in maternal serum

Stoops

Winget

DeChant

et al. 2018

Molecular Reproduction Devel

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Genetic sexing of animals with long gestation time benefits the management of captive populations. Here, X and Y chromosome-specific primers, based on equine gene sequencing data, were developed and tested on captive rhinoceroses (10 males, 20 females) representing four species (Diceros bicornis, Certaotherium simum simum, Rhinoceros unicornis, and Dicerorhinus sumatrensis). The Y chromosome-specific primer set targeted SRY (Sex-determining region Y), and amplified a 177-bp product following PCR of DNA extracted from males, but not females, of all species. A primer set based on the equine AMEL (Amelogenin) gene resulted in a 232-bp product following PCR of all rhinoceros species. These gene-specific primer sets were then evaluated for their ability to determine gender in cell-free DNA from rhinoceros serum. Modifications to the original extraction and PCR protocols were required to obtain sufficient DNA quantities from serum, and both DNA yield and PCR amplification were substantially reduced or absent following multiple freeze-thaw cycles of serum. When fresh serum from 14 pregnant rhinoceroses (ultimately bearing seven male and seven female calves), representing four species at different stages of gestation (Days 61-490), were probed in a PCR-based assay, an accuracy of 71% was achieved for male-specific gene detection of SRY, which improved to 100% by including a reamplification step into the protocol. Such early sex determination should be a valuable tool for current management practices as well as future assisted reproduction of rhinoceroses.

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Fluctuation of Maternal and Fetal Free Extracellular Circulatory DNA in Maternal Plasma

Cited by 22 publications

References 18 publications

Quantification of all fetal nucleated cells in maternal blood between the 18th and 22nd weeks of pregnancy using molecular cytogenetic techniques

Quantification of all fetal nucleated cells in maternal blood between the 18th and 22nd weeks of pregnancy using molecular cytogenetic techniques

Prenatal genotyping ofRHDandSRYusing maternal blood

Early fetal sexing in the rhinoceros by detection of male‐specific genes in maternal serum

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