Analysis of AmpFlSTR^® MiniFiler™ loci and its forensic application

Article Outline

• Abstract
• 1. Introduction
• 2. Materials and methods
• 3. Results and discussion
• Conflict of interest statement
• References
• Copyright

Abstract 

The allele frequencies of eight MiniFiler™ loci have been analyzed in 101 Japanese individuals living in Kanagawa with informed consent by means of ABI 310 Genetic Analyzer. A total of 7 alleles for D13S317, 8 alleles for D7S820, 11 alleles for D2S1338, 11 alleles for D21S11, 5 alleles for D16S539, 14 alleles for D18S51, 8 alleles for CSF1PO, and 13 alleles for FGA were observed. The polymorphic profiles of these MiniFiler™ loci in the present study were essentially the same as those obtained by using the AmpFlSTR^® Identifiler^® PCR Amplification kit. The combined matching probability of eight MiniFiler™ loci and cumulative probability of paternity exclusion were estimated as 1.97×10⁻¹⁰ and 0.9996, respectively. The MiniFiler™ kit was useful for individual identification in forensic analysis.

Keywords: DNA polymorphism, STR, Population study, Multiplex PCR amplification, Japanese population

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1. Introduction 

STR (short tandem repeat) polymorphisms are powerful tools for human identification, paternity analysis and genetic mapping [1]. The AmpFlSTR^® MiniFiler™ PCR Amplification kit was developed to recover information from DNA evidence, specifically degraded and/or inhibited samples. The MiniFiler™ kit was designed to amplify eight autosomal STR loci, D13S317, D7S820, D2S1338, D21S11, D16S539, D18S51, CSF1PO, and FGA, and the sex determining marker Amelogenin. In the present study, we analyzed these MiniFiler™ loci to obtain allele frequency data for a Japanese population living in Kanagawa and its forensic application was also examined.

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2. Materials and methods 

Blood samples were obtained from 101 unrelated healthy donors with informed consent. DNA was extracted using Quick Gene-800 (FUJIFILM, Japan). Multiplex PCR amplification was performed with MiniFiler™ kit according to the user's manual. The PCR products were electrophoresed by ABI 310 Genetic Analyzer and the profiles were evaluated using GeneMapper^® ID Software v3.2.

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3. Results and discussion 

In 101 Japanese subjects, the distribution of allele frequencies of eight MiniFiler™ loci, heterozygosities, polymorphism information contents (PIC), matching probabilities (pM) and probabilities of paternity exclusion (PE) were indicated in Table 1. A total of 7 alleles for D13S317, 8 alleles for D7S820, 11 alleles for D2S1338, 11 alleles for D21S11, 5 alleles for D16S539, 14 alleles for D18S51, 8 alleles for CSF1PO, and 13 alleles for FGA were observed. The distribution of genotypes of these eight MiniFiler™ loci fitted the Hardy–Weinberg equilibrium. The polymorphic profiles of these MiniFiler™ loci in the present study were essentially the same as those obtained by using the AmpFlSTR^® Identifiler^® PCR Amplification kit [2]. The combined matching probability of eight MiniFiler™ loci and cumulative probability of paternity exclusion were estimated as 1.97×10⁻¹⁰ and 0.9996, respectively. The combinational resolution of these eight STR systems has been shown to be a powerful tool in personal identification and paternity analysis. For the identification of skeletal remains, a postmortem paternity test was done with MiniFiler™ kit. An unequivocal conclusion was obtained. From the present study, the MiniFiler™ kit was applicable to routine case work.

Table 1. Analysis of Japanese population with MiniFiler™ loci.

	Locus
	D13S317	D7S820	D2S1338	D21S11	D16S539	D18S51	CSF1PO	FGA
Allele frequency (%)
Allele 7		0.49					0.99
Allele 8	23.27	13.37					0.49
Allele 9	13.37	5.94			31.68		3.47
Allele 10	11.88	26.24			19.80	0.49	19.31
Allele 11	24.75	27.23			23.27		19.31
Allele 12	21.78	20.79			16.34	3.96	47.03
Allele 13	4.45	4.95			8.91	19.31	6.93
Allele 14	0.50	0.99				22.28	2.47
Allele 15						17.82
Allele 16						11.88
Allele 17			10.40			5.94
Allele 18			16.34			8.42		4.95
Allele 19			21.78			3.47		5.94
Allele 20			9.90			1.48		9.90
Allele 21			0.99			0.99		11.39
Allele 22			5.94			1.98		20.79
Allele 22.2								0.99
Allele 23			11.88			0.99		21.29
Allele 23.2								1.48
Allele 24			8.91			0.99		12.38
Allele 25			7.92					6.44
Allele 25.2								0.49
Allele 26			2.47					2.97
Allele 27			3.47					0.99
Allele 28				3.47
Allele 28.2				0.49
Allele 29				29.70
Allele 30				33.17
Allele 30.2				0.49
Allele 31				7.92
Allele 31.2				4.46
Allele 32				2.48
Allele 32.2				14.85
Allele 33.2				1.98
Allele 34.2				0.99
Heterozygosity (%)	80.3	79.0	87.1	76.9	77.2	85.3	69.8	86.2
PIC	0.774	0.758	0.858	0.736	0.735	0.836	0.658	0.847
pM	0.068	0.076	0.030	0.086	0.089	0.038	0.131	0.034
PE	0.608	0.588	0.743	0.566	0.553	0.708	0.472	0.725

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Conflict of interest statement 

None.

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References 

1. Nakamura S, Nagai T, Matsui H, Tabuchi M, Sugie H, Furukawa M, et al. Genetic analysis of AmpFlSTR^® profiler plus loci in Japanese. Prog. Forensic Genet. 2000;8:236–238
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2. Yoshida K, Takahashi K, Kasai K. Allele frequencies of 15 loci using AmpFlSTR^® Identifiler Kit in Japanese population. J. Forensic Sci. 2005;50:718–719
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