Due to the high rate of complications, special medical care must be provided especially for monozygotic twin pregnancies, which are characterized as having 2.5 times higher mortality of fetuses. In recent years, examination of cell-free DNA (cfDNA) circulating in maternal plasma has become a useful noninvasive method of prenatal diagnosis. However, fetal DNA constitutes only 3–20% of plasma cfDNA during pregnancy. Short tandem repeats (STRs) are routinely used in forensic examination of DNA mixtures and are able to identify 5% minority components. Haplotypes of deletion/insertion polymorphisms and STRs (DIP–STRs) are able to detect even 0.1% minority components of DNA mixtures. Thus, STRs and DIP–STRs seem to be a perfect tool for detection of fetal alleles in DNA isolated from maternal plasma. Here, we present a novel noninvasive prenatal diagnosis technique of determination of pregnancy zygosity based on examination of feto-maternal microchimerism of plasma cfDNA with the use of STRs and DIP–STRs. Our preliminary results based on 22 STR loci showed 67% sensitivity, 100% specificity and 82% accuracy for prenatal detection of twin dizygosity. The corresponding values for seven DIP–STRs were 13%, 100% and 54%, respectively. Owing to assay performance, low DNA input requirements, low costs (below 10 USD per patient) and simplicity of analysis, genotyping of STR/DIP–STR markers in maternal plasma cfDNA may become a useful supplementary test for noninvasive prenatal diagnosis of twin zygosity in cases when chorionicity and zygosity cannot be reliably determined by ultrasound examination and prognostic value may be provided by a DNA test determining pregnancy zygosity.

This study reports on zygosity determination in twins of childhood age. Parents responded to questionnaire items dealing with twin similarity in physical characteristics and frequency of mistaking one twin for another by parents, relatives and strangers. The accuracy of zygosity diagnosis was evaluated across twins aged 6, 8, and 10 and across parents. In addition, it was examined whether the use of multiple raters and the use of longitudinal data lead to an improvement of zygosity assignment. Complete data on zygosity questions and on genetic markers or blood profiles were available for 618 twin pairs at the age of 6 years. The method used was predictive discriminant analyses. Agreement between zygosity assigned by the replies to the questions and zygosity determined by DNA markers/blood typing was around 93%. The accuracy of assignment remained constant across age and parents. Analyses of data provided by both parents and collected over multiple ages did not result in better prediction of zygosity. Details on the discriminant function are provided. Twin Research (2000) 3, 134–141.

The use of DNA restriction fragment length polymorphisms (RFLP) to analyze variable number of tandem repeat (VNTR) sequences dispersed in the human genome, has become a powerful tool for the study of population genetics due to the very substantial polymorphism involved. Because the markers usually employed for twin zygosity determination (such as sex combination, placentation, HLA typing, blood group antigens, etc) may not be uniformly informative, we propose the use of synthetic olygonucleotides, representing VNTR “core” sequences, for the determination of zygosity in twins.

The incidence of preterm delivery among twin maternities in Oxford between 1973 and 1983, was 39.4%. Preterm delivery was more common in MZ than DZ twins, and this was associated with higher incidence of spontaneous rupture of the membranes associated with monozygosity.

A statistical index is proposed in order to determine twin zygosity based on weight and height measures and a Bayesian test of hypotheses. Some Roman twin data are analyzed on the basis of the Euclidean distance index, which appears to be the best for an easy calculation and for its genetic interpretation based on the formulation of genetic distance.

A mailed questionnaire dealing with similarity in childhood, and how often and by whom the twins were mixed up, was applied to 290 same-sex adult twin pairs. For about 75% of pairs ten genetic markers were analysed. The agreement in zygosity classification between questionnaire and blood typing was high. Irrespective of whether raw scores or discriminant function analysis was applied, the agreement was nearly 95%. When a decision tree was applied, the percentage rose to nearly 96%. Considering that probably four of the twin pairs were wrongly classified as MZ by genetic markers, the percentage rose to 96 and 98, respectively.

The use of epidemiological studies of twins as a means of determining the existence of genetic factors in various conditions is well known. To be of value, the zygosity of the twin pairs must be identified with as great a degree of confidence as possible. A method of determining twin zygosity, using concordance in general likeness, hair colour, hair type, eye colour, ear form, tongue roll and phenylthiocarbamide (PTC) taste sensitivity, was developed for use in a survey of 244 pairs of like-sexed twins aged 5 to 15 years. The results obtained and the problems raised when applying this method are discussed. The method was simple to apply to a large group of twin pairs, and monozygotic pairs were identified with a degree of confidence greater than 95%.