title: An integrated system of ABO typing and multiplex STR testing for forensic DNA analysis.
abstract: A new amplification system for ABO and STR genotyping in a single reaction has been successfully developed. Two types of information can be obtained from a biological sample at one time. One is the classical information of ABO blood group typing for screening suspects and the other is STR information for individual identification. The system allows for the simultaneous detection of 15 autosomal STR loci (containing all CODIS STR loci as well as Penta D and Penta E), six ABO genotypes (O/O, B/B, A/A, A/O, A/B, and B/O) and the gender-determining locus Amelogenin. Primers are designed so that the amplicons are distributed ranging from 75bp to 500bp within a four-dye fluorescent design, leaving a fourth dye for the internal size standard. With 30 cycles, the results showed that the optimal amount of DNA template for this multiplex ranges from 250pg to 2ng and the lowest detection threshold is 125pg (as low as 63pg for ABO loci). For the DNA template outside the optimal detection range, we could adjust the number of cycles to obtain the robust profiles. Mixture studies showed that over 83% of minor alleles were detected at 1:9 ratios. The full profiles were still observed when 4ng of degraded DNA was digested by DNase I and 1ng undegraded DNA was added to 40μM haematin. Polymerase chain reaction (PCR)-based conditions including the concentrations of primers, magnesium and the Taq polymerase as well as volume, cycle numbers and annealing temperature were examined and optimised. In addition, the system was validated by 364 bloodstain samples and 32 common casework samples. According to the Chinese National Standards and Scientific Working Group on DNA Analysis Methods (SWGDAM) guidelines, our system demonstrates good detection performance and is an ideal tool for forensic DNA typing with potential application.
authors.affiliation: Criminal Science and Technology Institute of Liaoning Province, No. 2 Qishan Middle Road, Huanggu District, Shenyang 110032, China. jiangxianhua_2006@yahoo.com.cn
authors.forename: Xianhua
authors.initials: X
authors.lastname: Jiang
authors.forename: Juan
authors.initials: J
authors.lastname: He
authors.forename: Fei
authors.initials: F
authors.lastname: Jia
authors.forename: Hongying
authors.initials: H
authors.lastname: Shen
authors.forename: Jinling
authors.initials: J
authors.lastname: Zhao
authors.forename: Chuguang
authors.initials: C
authors.lastname: Chen
authors.forename: Liping
authors.initials: L
authors.lastname: Bai
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.forename: Guangwei
authors.initials: G
authors.lastname: Hou
authors.forename: Faye
authors.initials: F
authors.lastname: Guo
mesh_terms: D000017:ABO Blood-Group System / Q000235:genetics*; D053523:Amelogenin / Q000235:genetics; D000818:Animals; D001788:Blood Grouping and Crossmatching; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers; D019075:Electrophoresis, Capillary; D005260:Female; D005787:Gene Frequency; D005828:Genetics, Population; D005838:Genotype; D006801:Humans; D008297:Male; D018895:Microsatellite Repeats; D060885:Multiplex Polymerase Chain Reaction; D013045:Species Specificity
chemical_list: D000017:ABO Blood-Group System; D053523:Amelogenin; D017931:DNA Primers