• Users Online: 219
  • Home
  • Print this page
  • Email this page
Home About us Editorial board Ahead of print Current issue Search Archives Submit article Instructions Subscribe Contacts Login 


 
 Table of Contents  
ORIGINAL ARTICLE
Year : 2018  |  Volume : 4  |  Issue : 3  |  Page : 122-128

The validation of Goldeneye™ DNA ID 22NC kit and the genetic polymorphism of 21 short tandem repeat loci in the Chinese Hunan Han population


Department of Forensic Medicine, School of Basic Medical Sciences, Central South University, Changsha, China

Date of Web Publication28-Sep-2018

Correspondence Address:
Prof. Zha Lagabayila
No. 172, Tongzipo Road, Changsha, Hunan
China
Login to access the Email id

Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jfsm.jfsm_18_18

Rights and Permissions
  Abstract 


Goldeneye™ DNA ID 22NC Kit is a novel short tandem repeat (STR) genotyping system that investigate 20 non-CODIS loci (D4S2366, D6S477, D22GATA198B05, D15S659, D8S1132, D3S3045, D14S608, D17S1290, D3S1744, D2S441, D18S535, D13S325, D7S1517, D10S1435, D11S2368, D19S253, D1S1656, D7S3048, D10S148, and D5S2500), a CODIS locus (D3S1358), and a sex-determining locus amelogenin in one assay. In the present study, this STR genotyping system was validated according to the guidelines of “Validation Guidelines for DNA Analysis Methods (2016)” updated by the Scientific Working Group on DNA Analysis Methods. A series of tests, such as polymerase chain reaction-based studies, sensitivity, inhibitors, DNA mixture, species specificity, precision and accuracy evaluation, stutter percentage, and peak height ratio, was conducted. The genetic polymorphism of 21 STR loci that included in the 22NC system was also investigated in the Chinese Hunan Han population. The validation results demonstrated that Goldeneye™ DNA ID 22NC Kit is a robust and reliable identification assay as required for genotyping in kinship analysis and forensic investigation. The 21 STR loci in this kit also showed a high level of genetic polymorphism for the Hunan Han population. Therefore, it can be used for forensic applications and population studies.

Keywords: 22NC, genetic polymorphism, Hunan Han, short tandem repeat


How to cite this article:
Fu X, Sun S, Liu Y, He J, Cai J, Lagabayila Z. The validation of Goldeneye™ DNA ID 22NC kit and the genetic polymorphism of 21 short tandem repeat loci in the Chinese Hunan Han population. J Forensic Sci Med 2018;4:122-8

How to cite this URL:
Fu X, Sun S, Liu Y, He J, Cai J, Lagabayila Z. The validation of Goldeneye™ DNA ID 22NC kit and the genetic polymorphism of 21 short tandem repeat loci in the Chinese Hunan Han population. J Forensic Sci Med [serial online] 2018 [cited 2018 Dec 17];4:122-8. Available from: http://www.jfsmonline.com/text.asp?2018/4/3/122/242509




  Introduction Top


As the second generation of genetic markers, short tandem repeats (STR) have been widely used as a biological evidence for kinship analysis and forensic investigation. The extensive distribution and highly polymorphic characteristics make the STR detection precise and specific in personal identity and paternity testing. With the current polymerase chain reaction (PCR) and capillary electrophoresis (CE) technology, a number of STR markers can be included in a multiplex kit to provide more genetic information and get higher values of the power of discrimination (PD) or probability of exclusion (PE)(Goldeneye ® Technology Ltd., Beijing, China) is a new STR genotyping system that investigate 20 non-CODIS loci (D4S2366, D6S477, D22GATA198B05, D15S659, D8S1132, D3S3045, D14S608, D17S1290, D3S1744, D2S441, D18S535, D13S325, D7S1517, D10S1435, D11S2368, D19S253, D1S1656, D7S3048, D10S148, and D5S2500), a CODIS locus (D3S1358), and the amelogenin locus in one assay. The 21 autosomal STR markers are distributed, respectively, across different chromosomes, which can greatly reduce the linkage probability to accommodate various forensic cases. Previous population data reported that these markers presented an extremely high polymorphism in the Tibetan and Li nationality.[1],[2]

In the present study, the Goldeneye™ DNA ID 22NC kit was validated according to the guidelines of “Validation Guidelines for DNA Analysis Methods (2016)” updated by the Scientific Working Group on DNA Analysis Methods.[3] A series of tests, such as PCR-based studies, sensitivity, inhibitors, DNA mixture, species specificity, precision and accuracy evaluation, stutter percentage, and peak height ratio, was conducted. The genetic polymorphism of 21 STR loci included in the 22NC system was also investigated in the Chinese Hunan Han population.

The Han nationality is the largest and most widely distributed people among the 56 ethnic groups in China. The tribes of the Yan and Yellow Emperors are the main predecessors of the Han group. With thousands of years of coexistence with other groups, the Han population has formed relationships with different groups. Currently, the Han people are mainly distributed in the Songliao plain, Yangtze River Basin, Yellow River Basin, and Pearl River Basin. Hunan province is located in the middle reaches of Yangtze River in Central South China and is situated between 108° to 114° east longitude and 24°–30° north latitude. Ethnic groups such as the Han, Tujia, Miao, Yao, Dong, Bai, Hui, and 55 others live in this province. The Han population in Hunan is close to 59.15 million and accounts for approximately 90% of the total population.[4] In this study, 504 unrelated individuals of Hunan Han were investigated. The allele frequencies, Hardy–Weinberg equilibrium (HWE), forensic parameters, and population differentiation were analyzed for evaluating the genetic polymorphism of 22NC system in Han nationality.


  Materials and Methods Top


Samples preparation

Typical control DNA 9948 and 9947A (Goldeneye ® Technology Ltd., Beijing, China) were applied to polymerase chain reaction-based studies, sensitivity, stability, and DNA mixture studies. The animal blood collected from pig, dog, fish, mouse, rat, and monkey were used for specificity testing. Following written informed consent of all participants, a total of 504 blood samples (260 females and 244 males) were collected from unrelated healthy Hunan Han individuals for accuracy evaluation, stutter percentage peak height ratio statistics, and population genetic polymorphism investigation. Genomic DNA of each blood samples was isolated using the salting-out method,[5] quantitated by the NanoDrop 1000 spectrophotometer, and analyzed by NanoDrop 2.4.7c software (NanoDrop Technologies Inc., Wilmington, DE, USA). The current study was performed in the Department of Forensic Science (Central South University, China) and was approved by the Ethics Committee of the Third Xiangya Hospital of Central South University (Approval Code: 2018-S194).

Polymerase chain reaction amplification and capillary electrophoresis

PCR was performed on a GeneAmp PCR 9700 thermal cycler (Thermo Fisher Scientific, Waltham, MA, USA). Except the tests in PCR-based studies, the experiment procedures were done strictly according to the manufacturer's instruction. The amplification reactions were 10 μL in total: 2.0 μL of primer mix, 2.0 μL of reaction mix, 5.0 μL of deionized water, and 1.0 μL of template DNA. Thermal cycling was conducted under the following conditions: an initial denaturation step at 95 °5 for 2 min, then 30 cycles of 94°C for 5 s, 60°C for 1 min, and finally an extension step at 60°C for 20 s. The amplification products were separated and detected by CE on the Applied Biosystems 3130 Genetic Analyzer (Applied Biosystems, Foster City, USA). About 1 μL of the PCR products or allelic ladder was combined with 9 μL of a 20:1 mixture of Hi-Di formamide (Applied Biosystems, Foster City, CA, USA) and ORG500 size standard for electrophoresis. Samples were injected at 3 kV for 10 s and electrophoresed with the run module of default parameters of HID Fragment Analysis36_POP4 and the filter set of G5 (Applied Biosystems, Foster City, CA). Genotyping was performed on the GeneMapper ID version 3.2 software (Applied Biosystems, Foster City, CA, USA). Unless otherwise stated, allele peaks were interpreted with a threshold of 50 RFU.

Polymerase chain reaction-based studies

The PCR key parameters were studied to evaluate the range of amplification conditions which could obtain the accurate STR genotyping results. The reaction volumes (5, 10, and 20 μL), primer mix concentration (1, 2, 4, and 8 μL), reaction mix concentration (1, 2, 4, and 8 μL), and annealing temperature (56°C, 58°C, 60°C, 62°C, and 64 °4) were tested in triplicate. Only tested parameter changed while others keep the same with the recommended conditions in a variety of conditions. The same concentration of 9948 DNA (1 ng/μL) was tested for reaction volume testing.

Sensitivity and stability studies

The 9948 DNA was diluted to evaluate the sensitivity of Goldeneye 22NC system. A serial 9948 dilutions with the DNA concentrations of 1, 0.5, 0.25, 0.125, 0.0625, and 0.0313 ng/μL were amplified in triplicate. Amplification and detection processes were performed following the manufacturer's recommended conditions. Percentage profile and mean peak height were determined for each template amount. The homozygotes were counted as one in the percentage profile. Two kinds of chemicals associated with common contaminants of casework samples, hematin, and humic acid were used in stability studies. 1 ng/μL of 9948 DNA was used for amplification while amplification reactions contained different concentrations of hematin (10, 20, 30, 40, and 50 μM) and humic acid (10, 20, 30, 40, and 50 ng/μL). Each test was performed three times.

Mixture and species specificity studies

To assess the ability of reliable detection of DNA mixtures with Goldeneye 22NC system, a mixed male/female DNA samples (9948 and 9947A) were examined at various ratios (19:1, 9:1, 4:1, 2:1, and 1:1) with 1 ng of total DNA template, and the test was performed three times to ensure the accuracy. For species specificity tests, genomic DNA of each animal (pig, dog, fish, mouse, and rat) was analyzed in triplicate with the use of Goldeneye 22NC system. These genomic DNA have been quantitated by the NanoDrop 1000 spectrophotometer to make sure the amount of DNA for each analysis was more than 1 ng.

Sizing precision and accuracy studies

Precision was measured by running the CE of the 18 allelic ladder samples on the 3130 Genetic Analyzer. For sizing accuracy, 50 random samples of Hunan Han nationality were genotyped under standard conditions, and the allele size was compared to the corresponding allelic ladder.

Stutter percentage and peak height ratio studies

For the stutter percentage and peak height ratio study, we had selected 300 samples randomly from the 504 blood samples. The threshold of minimum stutter peak height was set to 10 RFU. Only peaks regarded at ± 1 repeat unit stutter position were included in the analysis. Stutter peaks at ± stutter position (in between two true allelic peaks) were excluded from the analysis. The percentage of stutter peaks of each 21 STR loci was determined by calculating the stutter peak height as a percentage of the main allele peak height from the 300 random samples. These 300 random samples were also used for the peak height ratio (PHR) calculation.

Population genetic polymorphism studies

We strictly adhered to the recommendations of the International Society for Forensic Genetics for the analysis of DNA polymorphisms and nomenclature.[6] Typical control DNA 9948 was used as a positive control and deionized water as a negative control during the population polymorphism analysis. Allele frequencies, exact tests of the HWE, and forensic parameters, including observed heterozygosity (OH), matching probability (MP), PD, PE, and typical paternity index (TPI), were evaluated using a modified PowerStat version 1.2 spreadsheet (Promega, Madison, WI, USA).[7] The exact test of population differentiation between the Hunan Han group and other previously published population data was analyzed with the Arlequin Version 3.5 software (Computationaland Molecular Population Gentics Lab, Zoological Institute, University of Berne, Bern, Switzerland).[8]


  Results and Discussion Top


Polymerase chain reaction-based studies

The recommended reaction volume for Goldeneye 22NC system is 10 μL. In this study, different reaction volumes (5, 10, and 20 μL) were tested in which the component ratio remained the same with the manufacturers' instruction. The genotyping profiles of 5 and 20 μL reaction volumes showed a high degree of consistency with recommended 10 μL reaction volume [Figure S1]. None allele dropped out. However, the peak height got from both reaction volumes of 5 and 20 μL was lower than 10 μL. The results suggested that the Goldeneye 22NC system could obtain correct genotypes when changed to halved or doubled standard reaction volume. When the technicians deal with low-copy number (LCN) DNA samples in actual cases, a smaller reaction volumes of this system could be choose to meet the requirements.[9]



Primer concentration may influence specificity and sensitivity of the STR analysis. The recommended concentration of the primer mix for Goldeneye 22NC system is 2 μL per 10 μL reaction volume. In the present study, different concentrations of primer mix (1, 2, 4, and 8 μL) were tested. None allele dropped out in the final electropherogram of each test [Figure S2]. Profiles of 1 μL primer mix were observed average 32.5% (13 of total 40 alleles) peak heights between 50 RFU and 200 RFU, significantly more than other groups. Both 4 and 8 μL of primer mix concentrations could provide sensitive and balanced amplification results. These tests suggested that even quadruple concentrations of primer mix would not have much impact on genotyping results.



The reaction mix contains the PCR Buffer, Mg2+, dNTPs, and Taq DNA polymerase. The recommended concentration of the reaction mix for Goldeneye 22NC system is 2 μL per 10 μL reaction volume. In the present study, different concentrations of reaction mix (1, 2, 4 and 8 μL) were tested. Correct genotyping results just obtained from the test of 2 μL reaction mix [Figure S3]. In the testing of 4 μL reaction mix, allele dropped out in D8S1132 and D3S1744, and the profiles were observed average 21.62% (8 of total 37 alleles) peak heights between 50 and 200 RFU. We could not detected the full profile from the other groups (1 and 8 μL of the reaction mix) either. These testings suggested that the concentration of contents in reaction mix has much impact on genotyping results. The volume of reaction mix should be carried out strictly according to the manufacturers' requirements.



The recommended annealing temperature for Goldeneye 22NC system is 60°C. In the present study, complete profiles generated and no allele dropout observed when annealing temperature ranged from 56°C to 62°C [Figure S4]. The allele peak heights slightly increased as annealing temperature from 56°C to 60°C, then reduced as temperature come up to 62°C. The profiles at 58°C and 62°C worked almost equally well on peak heights and peak height balance with the recommended 60°C. Hence, 58°C to 62°C, all could be the appropriate annealing temperature. Alleles of D4S2366, D15S659, D8S1132, AMEL, D18S535, and D19S253 locus dropped out when annealing temperature turned to 64°C. These results confirmed Goldeneye 22NC system was adapting well to differential annealing temperatures, which can reduce the possibility of unsuccessful amplification due to inaccurate temperature.



Sensitivity and stability studies

Different quality of DNA samples would be obtained from the forensic investigations, some of which were beyond the manufacturers' requirements. LCN DNA and inhibitors such as hematin and humic acid are common in the actual case samples. The sensitivity and stability study can determine the limits of sample DNA quantities and the influence of existing inhibitors.

In sensitivity study, full profiles were obtained using 50 RFU peak heights analysis threshold with the concentration of 9948 above 0.25 ng [Figure S5]. As template DNA gradually reducing from 1 to 0.25 ng, the average peak height was detected from 1102 to 262 RFU. When the template DNA down to 0.125 ng, partial profiles were observed, and average 20% (8 of total 40 alleles) of the loci were detected with the average peak height at 123 RFU. When the template DNA down to 0.0625 ng, 12% (5 of total 40 alleles) of the loci were detected with the average peak height at 58 RFU. All the loci dropped out when template DNA reduced to 0.0313 ng [Figure 1]. These results suggested that the sensitivity of Goldeneye 22NC system was determined to be 0.25 ng in 10 μL PCR reaction volume.

Figure 1: Sensitivity testing of template DNA ranged from 1 to 0.0313 ng

Click here to view


Humic acid was thought to affect the template availability for amplification by binding to DNA, and hematin could curb the function of Taq polymerase.[10] In this study, different concentrations of hematin (10, 20, 30, 40, and 50 μM) and humic acid (10, 20, 30, 40, and 50 ng/μL) were used as common inhibitors in stability studies. Full profiles were observed with hematin up to 50 μM [Figure S6] and humic acid concentration up to 40 ng/μL [Figure S7]. The profile average peak heights were decreasing gradually when the inhibitors concentration was increasing. When the humic acid concentration up to 50 ng/μL, the allelic peak dropped out in D6S477 with the profile average peak heights at 763 RFU.



Mixture and species specificity studies

Samples mixed by other individual or nonhuman DNA appeared frequently in forensic casework. To assess the ability of reliable detection of mixtures with Goldeneye 22NC system, a mixed male/female (9948 and 9947A) DNA samples were examined at various ratios (1:1, 2:1, 4:1, 9:1, and 19:1). As the proportion of minor contributor DNA 9947A reducing, a concomitant reducing in peak height was observed for minor alleles [Figure S8]. All of the minor alleles were detected for ratios of 1:1, 2:1, and 4:1. When at 9:1 ratio of 9948 and 9947A, several alleles (average 22.5%) of minor contributor dropped out, and some alleles were confused with stutter peaks. Things got much worse at 19:1 ratio with more minor alleles (average 40%) dropped out [Figure 2]. These results suggested that the Goldeneye 22NC system was able to deal with a DNA mixture confined to the ratio of 4:1.

Figure 2: A total of 1 ng DNA template mixed by 9948 and 9947A at various ratio (1:1, 2:1, 4:1, 9:1, and 19:1) were tested in triplicate

Click here to view


The common animals (pig, dog, fish, mouse, rat, and monkey) which were closely related to human activity were analyzed with the Goldeneye 22NC system for species specificity. Peaks above 50 RFU were detected from pig, dog, fish, and monkey. For pig, an “OL” peak (peak height 1326 RFU) at amelogenin locus with size 92.44 bp and an “OL” peak (peak height 540 RFU) at D18S535 with size 123.29 bp were detected. These peaks were out of panels and definitely had no material effect on genotyping results. For dog, an allele of “13” (peak height 120 RFU) at D4S2366 locus with size 101.95 bp and an allele of “19” (peak height 102 RFU) at D3S1358 locus with size 150.03 bp was detected. For fish, an allele of “12” (peak height 136 RFU) at D4S2366 locus with size 102.01 bp, an allele of “17” (peak height 65 RFU) at GATA198B05 locus with size 221.04 bp, an allele of “X” (peak height 64 RFU) at the amelogenin locus with size 95.96 bp, an allele of “19” (peak height 108 RFU) at D3S1358 locus with size 150.17 bp, and loci of “13, 15” (peak height 89 RFU and 115 RFU, respectively) at D2S441 locus with size 95.77 bp and 103.64 bp were detected. These peaks were in low peak height and hardly effected on genotyping results. For monkey, lots of peaks (peak height from 89 to 6939 RFU), including “OL” and amelogenin allele, were observed with the size ranged from 71.74 to 358.02 bp, these peaks would affect genotyping to some extent [Figure S9]. The species specificity results suggested that it should be cautious for samples possibly combined with DNA from monkey.



Sizing precision and accuracy studies

Sizing precision study is of great significance to accurate and reliable genotyping. 18 allelic ladder samples were detected on the 3130 Genetic Analyzer. The average size and standard deviation for each allele were computed and showed to be from 0.0216 (D13S325) to 0.1756 (D4S2366) [Figure 3]. The average standard deviation of 18 allelic ladders was 0.0987, demonstrated the system had good precision.
Figure 3: Size variation of all allelic ladders performed on 3130 Genetic Analyzer (n = 18)

Click here to view


The sizing accuracy was tested by computing the size discrepancy between all the STR alleles from 50 random samples and the corresponding allelic ladder. All sample alleles (total of 1978 alleles from 50 samples) were sized within ± 0.5 bp of a corresponding allele, indicated that there had a poor chance of genotyping mistake owing to insufficient sizing accuracy.

Stutter percentage and peak height ratio studies

Stutter peaks often manifest as one repeat unit longer or shorter than the main allele peak. They were caused by strand slippage during PCR amplification.[11] A high percentage of stutter may make it more difficult to analysis mixtures due to confusion with the minor contributor.[12] Stutter percentage and its relative allele peak heights for each STR were obtained from the analysis of 300 random samples of Hunan Han nationality with Goldeneye 22NC system. The results of stutter percentage and standard deviations are listed in [Table 1]. The lowest average percentage of stutter was observed at locus D17S1290 (5.06%), and the highest was D7S1517 (12.03%). Compared with the recommended stutter filter values, D14S608, D15S659, and D1S1656 had the stutter peaks in excess of the labeled line. The recommended stutter filters could contribute to analysis the genotyping of mixed samples, and maybe a fine complement to existing STR profiling capabilities.
Table 1: Stutter percentages were calculated for the 21 short tandem repeats loci from 300 samples

Click here to view


The PHR levels revealed the balance of heterozygotes. A good balance in all analyzed markers satisfied the requirements of correct and reliable genotyping. In this study, the calculation of the PHR was performed by analyzing 300 individuals obtaining an average PHR of 83.86%. It is notable that all of the loci in this study were observed PHR averages over 80% and the lowest PHR average loci were D2S441 (80.89%). Significantly, the loci of amelogenin were observed PHR averages over 90% (92.20%) and showed 1.48% PHR lower than 70% [Table 2]. Because several loci have the low PHR values, it is best to repeat the tests three times when deal with LCN DNA to avoid missing alleles. In brief, the Goldeneye 22NC system provided a decent balance of heterozygous alleles.
Table 2: Peak height ratio values were computed of 22 short tandem repeats loci

Click here to view


Population genetic polymorphism studies

In the present study, the 22NC system showed great detection efficiency and the electropherograms of all samples were obtained. A total of 276 alleles and 1125 genotypes were observed. The allele frequencies and forensic parameters of the 21 STR loci in Hunan Han population are shown in [Table S1]. The corresponding allelic frequencies ranged from 0.0010 to 0.3658. All 21 STR loci were exhibited high genetic polymorphism in the Hunan Han population, and the unique allele from these markers was between 9 (D3S3045 locus and D19S253) and 18 (D7S1517 locus). The values of the OH ranged from 0.6329 (D6S477 locus) to 0.9345 (D7S3048 locus). The observed MP, TPI, and PIC ranged from 0.0182 (D22GATA198B05 locus) to 0.1064 (D3S1358 locus), 1.3622 (D6S477 locus) to 7.6364 (D7S3048 locus), and 0.7101 (D3S1358 locus) to 0.9084 (D22GATA198B05 locus), respectively. Deviations from the HWE were detected at D19S253 (P = 0.0043). However, after the Bonferroni correction assuming that the 0.05 significance level obtained for the 21 tests (one per locus) yields an actual significance threshold of 0.0024,[13] no loci were against the conditions of the HWE. The PD ranged from 0.8936 (D3S1358 locus) to 0.9818 (D22GATA198B05 locus), and the combined PD was 0.999999999999999999999999992. The power of exclusion ranged from 0.3323 (D6S477 locus) to 0.8663 (D7S3048 locus), and the combined power of exclusion was 0.999999998.



The allelic frequencies of the 21 STR loci in the present study were compared to previously published population data from other groups in China. Not all the 21 STR loci were compared because of the limitations of previously published population data. In general, Eastern Han at 21 STR loci,[14] Tibetan at 21 STR loci,[1] Hainan Li at 21 STR loci,[2] Northern Han at 18 STR loci,[15] Uygur at 18 STR loci,[16],[17] Southern Han at 17 STR loci,[18],[19] Western Han at 17 STR loci,[20],[21] and Mongolian at 6 STR loci [22],[23] were compared with the Hunan Han group, and the P values are shown in [Table S2]. After applying the Bonferroni correction, the compared AMOVA results showed statistically significant differences (P < 0.0024) at 16, 15, 13, 12, 12, 8, and 5 STR loci between the Hunan Han and the Uygur, Northern Han, Southern Han, Eastern Han, Tibetan, Western Han, and Mongolian populations, respectively. The STR loci of D19S253, D6S477, D22GATA198B05, D15S659, D8S1132, D3S1358, D2S441, D18S535, D4S2366, D1S1656, and D7S3048 showed higher population differentiation than the other loci in the panel. These comparisons confirmed that allelic frequency distributions varied in different ethnic groups and in the same ethnic group living in different regions.




  Conclusion Top


The Goldeneye™ DNA ID 22NC Kit was developed to meet the demands for complex kinship analysis and forensic investigation. This kit investigated 20 non-CODIS loci (D4S2366, D6S477, D22GATA198B05, D15S659, D8S1132, D3S3045, D14S608, D17S1290, D3S1744, D2S441, D18S535, D13S325, D7S1517, D10S1435, D11S2368, D19S253, D1S1656, D7S3048, D10S148, and D5S2500), one CODIS locus (D3S1358), and the amelogenin locus with a 5-dye fluorescent analysis system. Previously reported data demonstrated that these markers presented an extremely high polymorphism in several populations. In the present study, we investigated the allelic frequencies and forensic data of 21 STR loci in the Hunan Han group through the Goldeneye™ DNA ID 22NC Kit. The STR loci belong to the 22NC system showed high genetic polymorphism in the Hunan Han population. The population data are valuable for the differentiation of individuals and the parentage testing of the Hunan Han people. In addition, the experiments confirmed the optimal reaction conditions and reagent concentrations for the amplification. The validation of the Goldeneye 22NC system was carried out and demonstrated that this commercial kit was robust, sensitive, and accurate. The system was suitable for personal identification and parentage testing especially when used in conjunction with other common STR kits.

Financial support and sponsorship

This study was financially supported by the National Natural Science Foundation of China (No. 81302615, 81302621, and 81571855).

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Gou H, Zhou J, Zhang Y, Wang C, Ma X, Ma H, et al. Allele frequency distribution of 21 forensic autosomal STR loci of goldeneye™ DNA ID 22NC kit in Chinese Tibetan Group. Forensic Sci Int Genet 2016;22:e21-4.  Back to cited text no. 1
    
2.
Fu X, Liu Y, Wang Z, Guo Y, Cai J, Zha L, et al. Genetic polymorphisms of 21 STR loci in goldeneye™ DNA ID 22NC kit for Chinese li Ethnic Group. Forensic Sci Int Genet 2017;28:e45-6.  Back to cited text no. 2
    
3.
Scientific Working Group on DNA Analysis Methods (SWGDAM), Validation Guidelines for DNA Analysis Methods. Available from: https://docs.wixstatic.com/ugd/4344b0_eca4ba88ecea4623b0e 17fdbed8c2b29.pdf. [Last accessed 2012 Dec 05].  Back to cited text no. 3
    
4.
Population Census Office under the State Council Department of Population and Employment Statistics National Bureau of Statistics Tabulation of the 2010 Population Census of the People's Republic of China. Beijing: China Statistics Press; 2012.  Back to cited text no. 4
    
5.
Miller SA, Dykes DD, Polesky HF. A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res 1988;16:1215.  Back to cited text no. 5
    
6.
Olaisen B, Bär W, Brinkmann B, Budowle B, Carracedo A, Gill P, et al. DNA recommendations 1997 of the International Society for Forensic Genetics. Vox Sang 1998;74:61-3.  Back to cited text no. 6
    
7.
Tereba A. Tools for analysis of population statistics. Profiles DNA 1999;3:14-6.  Back to cited text no. 7
    
8.
Excoffier L, Lischer HE. Arlequin suite ver 3.5: A new series of programs to perform population genetics analyses under linux and windows. Mol Ecol Resour 2010;10:564-7.  Back to cited text no. 8
    
9.
Zhang S, Bian Y, Tian H, Wang Z, Hu Z, Li C, et al. Development and validation of a new STR 25-plex typing system. Forensic Sci Int Genet 2015;17:61-9.  Back to cited text no. 9
    
10.
Opel KL, Chung D, McCord BR. A study of PCR inhibition mechanisms using real time PCR. J Forensic Sci 2010;55:25-33.  Back to cited text no. 10
    
11.
Schlötterer C, Tautz D. Slippage synthesis of simple sequence DNA. Nucleic Acids Res 1992;20:211-5.  Back to cited text no. 11
    
12.
Walsh PS, Fildes NJ, Reynolds R. Sequence analysis and characterization of stutter products at the tetranucleotide repeat locus vWA. Nucleic Acids Res 1996;24:2807-12.  Back to cited text no. 12
    
13.
Weir BS. Genetic Data Analysis II: Methods for Discrete Population Genetic Data. Sunderland: Sinauer Associates; 1996.  Back to cited text no. 13
    
14.
Lin Y, Que TZ, Zhao ZM, Zhang SH. Forensic investigation of goldeneye™ DNA ID 22NC kit. Fa Yi Xue Za Zhi 2015;31:280-3.  Back to cited text no. 14
    
15.
Xie B, Chen L, Yang Y, Lv Y, Chen J, Shi Y, et al. Genetic distribution of 39 STR loci in 1027 unrelated Han individuals from Northern China. Forensic Sci Int Genet 2015;19:205-6.  Back to cited text no. 15
    
16.
Song F, Li J, Zhang L, Luo H. Population data for 22 autosomal STR loci in the Uygur ethnic minority. Int J Legal Med 2017;131:651-2.  Back to cited text no. 16
    
17.
Yuan L, Liu H, Liao Q, Xu X, Chen W, Hao S, et al. Genetics analysis of 38 STR loci in Uygur population from Southern Xinjiang of China. Int J Legal Med 2016;130:687-8.  Back to cited text no. 17
    
18.
Chen L, Lu H, Qiu P, Yang X, Liu C. Polymorphism analysis of 15 STR loci in a large sample of Guangdong (Southern China) Han population. Leg Med (Tokyo) 2015;17:489-92.  Back to cited text no. 18
    
19.
Liu QL, Chen ZX, Chen CG, Lu DJ. Genetic polymorphism of 22 autosomal STR markers in a Han population of Southern China. Forensic Sci Int Genet 2016;24:e14-e16.  Back to cited text no. 19
    
20.
Wang J, Hu XJ, Song XB, Zhou Y, Li SY, Zhou J, et al. Allele frequencies of nine non-CODIS STR loci in Western Chinese Han population. Forensic Sci Int Genet 2013;7:e88-9.  Back to cited text no. 20
    
21.
Li J, Luo H, Song F, Hou Y. Genetic polymorphisms of 22 autosomal STR loci in Chinese Han population. Forensic Sci Int Genet Suppl Ser 2015;5:e45-7.  Back to cited text no. 21
    
22.
Zha L, Liu Y, Guo Y, Li J, Wang K, Geng K, et al. Genetic polymorphism of 21 non-CODIS STR loci in the Chinese Mongolian ethnic minority. Forensic Sci Int Genet 2014;9:e32-3.  Back to cited text no. 22
    
23.
Li DX, Bi LF, Su XL. Genetic polymorphism of 6 short tandem repeat loci in mongolian population of China. Zhonghua Yi Xue Yi Chuan Xue Za Zhi 2004;21:407-9.  Back to cited text no. 23
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2]



 

Top
 
 
  Search
 
Similar in PUBMED
   Search Pubmed for
   Search in Google Scholar for
 Related articles
Access Statistics
Email Alert *
Add to My List *
* Registration required (free)

 
  In this article
Abstract
Introduction
Materials and Me...
Results and Disc...
Conclusion
References
Article Figures
Article Tables

 Article Access Statistics
    Viewed575    
    Printed58    
    Emailed0    
    PDF Downloaded77    
    Comments [Add]    

Recommend this journal


[TAG2]
[TAG3]
[TAG4]