Genetic testing kits and a method of bladder cancer
View Patent ↗A deletion in the end region of the long arm of a Chromosome 9 is efficiently detected. A genetic testing kit of bladder cancer according to the present invention includes a primer allowing for efficient amplification of a region containing a site of genetic polymorphism present in the ABO blood group gene of Chromosome 9. In the site of genetic polymorphism present in the ABO blood group gene, the frequency of heterozygote (heterozygosity) in the population is extremely high. Therefore, by detecting LOH using a polymorphic site present in the ABO blood group gene, it is possible to reliably detect a deletion near the polymorphic site, in other words, a deletion near the end of the long arm of Chromosome 9.
1. A method for detecting loss of heterozygosity in human, comprising analyzing a biological sample of a human subject for a polymorphic genotype or genotypes at, at least, one of the 261st and the 297th sites of the sequence of SEQ ID NO:4 located in the ABO blood group gene on Chromosome 9, and detecting loss of heterozygosity at said site or sites based on the presence of heterozygosity of the results of the polymorphism analysis, wherein the polymorphic genotype at the 261st is guanine or deletion of guanine and the polymorphic genotype at the 297th site is guanine or adenine, and wherein the detection of the loss of heterozygosity is indicative of, at least, a partial deletion of Chromosome 9.
2. The method according to claim 1 , wherein the analysis is performed by amplifying a nucleic acid region containing said site or sites of polymorphism.
3. The method according to claim 2 , wherein the amplification is performed by using a first primer containing 5 or more continuous bases of the base sequence 5′-TCTCCATGTGCAGTAGGAAAGGATG-3′ (SEQ ID NO:1) and a second primer containing 5 or more continuous bases of the base sequence 5′-CACAGTTAACCCAATGGTGGTGTT-3′ (SEQ ID NO:2).
4. The method according to claim 1 , wherein the loss of heterozygosity is detected by a single-strand conformation polymorphism analysis.
5. The method according to claim 4 , wherein single-strand conformation polymorphism analysis is performed by capillary electrophoresis.
6. The method of claim 1 further comprising analyzing the biological sample for a polymorphic genotype or genotypes at, at least, one additional site selected from the group consisting of D9S304, D9S303, ALDOB, and D9S775, and detecting loss of heterozygosity at said site or sites based on the presence of heterozygosity of the results of the polymorphism analysis, wherein the loss of heterozygosity at any of said sites is indicative of, at least, a partial deletion of Chromosome 9.
7. The method of claim 6 , wherein the analysis is performed by amplifying a nucleic acid region containing the site or sites of polymorphism.