The TDI-FP assay in human Y chromosome SNP haplotyping.
One of the many commercial technologies for genotyping single nucleotide polymorphisms (SNPs) is template direct dye-terminator incorporation with fluorescence-polarization (TDI-FP assay). It is a single-base extension assay followed by reading the fluorescence polarization values in an appropriate instrument. We have evaluated the suitability of the TDI-FP technique to detect haploid uniparentally inherited DNA polymorphisms on the nonrecombining portion of the Y chromosome. A sample of 47 individuals has been genotyped for 8 Y chromosome biallelic markers. The SNP typing was blindly duplicated by the denaturing high-performance liquid chromatography (DHPLC) technique for comparison. In the cases under examination the TDI-FP assay was able to resolve an allelic state fully. Such a result showed 100% concordance indicating how efficiently the TDI assay can be used to genotype Y chromosome DNA SNPs. However, a percentage of indeterminate genotypes remained unresolved by simple visual inspection: it varied from 0% to 11% depending on the SNP locus and on the success of amplification. This is consistent with previous findings. A maximum likelihood classificatory analysis allowed some of the indeterminate genotypes to be assigned and some potentially misclassified samples to be identified. Their percentage remains relatively high despite retyping and therefore alternative techniques for these noncompliant situations are required.
Di Gaetano
C
C
Dipartimento di Genetica, Biologia e Biochimica, Università di Torino, 10126 Torino, Italy.
Crobu
F
F
Guarrera
S
S
Polidoro
S
S
Gasparini
M
M
Underhill
P A
PA
Matullo
G
G
Piazza
A
A
D001331:Automation; D002851:Chromatography, High Pressure Liquid; D041322:Chromosomes, Human, Y; D017931:DNA Primers; D005454:Fluorescence Polarization; D005838:Genotype; D006239:Haplotypes; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D013728:Terminator Regions, Genetic
D017931:DNA Primers