| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Received on May 27, 2008
Accepted on July 25, 2008
Molecular Diagnostics and Genetics |
1 Centre for Research into Circulating Fetal Nucleic Acids, Li Ka Shing Institute of Health Sciences, and Department of Chemical Pathology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong
2 Department of Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong
* To whom correspondence should be addressed. E-mail: loym{at}cuhk.edu.hk.
BACKGROUND: The precise measurement of cell-free fetal DNA in maternal plasma facilitates noninvasive prenatal diagnosis of fetal chromosomal aneuploidies and other applications. We tested the hypothesis that microfluidics digital PCR, in which individual fetal-DNA molecules are counted, could enhance the precision of measuring circulating fetal DNA.
METHODS: We first determined whether microfluidics digital PCR, real-time PCR, and mass spectrometry produced different estimates of male-DNA concentrations in artificial mixtures of male and female DNA. We then focused on comparing the imprecision of microfluidics digital PCR with that of a well-established nondigital PCR assay for measuring male fetal DNA in maternal plasma.
RESULTS: Of the tested platforms, microfluidics digital PCR demonstrated the least quantitative bias for measuring the fractional concentration of male DNA. This assay had a lower imprecision and higher clinical sensitivity compared with nondigital real-time PCR. With the ZFY/ZFX assay on the microfluidics digital PCR platform, the median fractional concentration of fetal DNA in maternal plasma was 
2 times higher for all 3 trimesters of pregnancy than previously reported.
CONCLUSIONS: Microfluidics digital PCR represents an improvement over previous methods for quantifying fetal DNA in maternal plasma, enabling diagnostic and research applications requiring precise quantification. This approach may also impact other diagnostic applications of plasma nucleic acids, e.g., in oncology and transplantation.
The following articles in journals at HighWire Press have cited this article:
|
|
 |
|
  C. F Wright and L. S Chitty Cell-free fetal DNA and RNA in maternal blood: implications for safer antenatal testing BMJ, July 6, 2009; 339(jul06_2): b2451 - b2451. [Full Text]
|
|
|
|
 |
|
 T. Chu, K. Bunce, W. A. Hogge, and D. G. Peters Statistical model for whole genome sequencing and its application to minimally invasive diagnosis of fetal genetic disease Bioinformatics, May 15, 2009; 25(10): 1244 - 1250. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E C W Hung, R W K Chiu, and Y M D Lo Detection of circulating fetal nucleic acids: a review of methods and applications J. Clin. Pathol., April 1, 2009; 62(4): 308 - 313. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. K.F. Yung, K.C. A. Chan, T. S.K. Mok, J. Tong, K.-F. To, and Y.M. D. Lo Single-Molecule Detection of Epidermal Growth Factor Receptor Mutations in Plasma by Microfluidics Digital PCR in Non-Small Cell Lung Cancer Patients Clin. Cancer Res., March 15, 2009; 15(6): 2076 - 2084. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R. W. K. Chiu, K. C. A. Chan, Y. Gao, V. Y. M. Lau, W. Zheng, T. Y. Leung, C. H. F. Foo, B. Xie, N. B. Y. Tsui, F. M. F. Lun, et al. Noninvasive prenatal diagnosis of fetal chromosomal aneuploidy by massively parallel genomic sequencing of DNA in maternal plasma PNAS, December 23, 2008; 105(51): 20458 - 20463. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
F. M. F. Lun, N. B. Y. Tsui, K. C. A. Chan, T. Y. Leung, T. K. Lau, P. Charoenkwan, K. C. K. Chow, W. Y. W. Lo, C. Wanapirak, T. Sanguansermsri, et al. Noninvasive prenatal diagnosis of monogenic diseases by digital size selection and relative mutation dosage on DNA in maternal plasma PNAS, December 16, 2008; 105(50): 19920 - 19925. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH |
