THE APPLICATION OF CELL-FREE FETAL DNA (cff-DNA) AND SIBLINGS DNA METHODS IN THE PROCESS OF PATERNITY TEST THROUGH CODIS STR LOCI (CSF1PO, THO1, TPOX, AND vWA)

Background: The non-invasive cff-DNA and siblings DNA methods are the latest breakthroughs in the forensic identification process. The use of cff-DNA and siblings DNA as non-invasive techniques in the forensic identification process has, hitherto, not been widely proven. Methods and Materials: This was an analytic observational study. The sample of this study consisted of peripheral blood of women in the second trimester of pregnancy and their two biological children. The kinship analysis was carried out through siblings' DNA and cff-DNA from the mothers through CODIS STR loci (CSF1PO, THO1, TPOX, and vWA). Results: The means of allele sharing between full siblings in loci CSF1PO, THO1, TPOX, and vWA were 0 (13.75%), 1 (44.75%), and 2 (41.50%). The allele sharing found in the study is in line with the one in previous research conducted by Wenk (1998) and the theory proposed by O'Connor (2011), indicating that one allele sharing dominates, contrasting with the finding of previous research conducted by Sosiawan (2020) revealing that 2-allele sharing was more superior. The variation is caused by the ethnicity having a different genetic contribution among the population. The variation can be attributed to historical and demographical processes leading to genetic drift. Conclusion: The mean of SI in 1 allele sharing in CODIS STR loci (CSF1PO, THO1, TPOX, and vWA) has the highest value of 44.5%. The use of cff-DNA of pregnant women as one of the non-invasive techniques can serve as an alternative material in a paternity test.

Potential Use of Cell Free Fetal DNA at 13 Short Tandem Repeats Loci for Noninvasive Prenatal Paternity Test

Background : Prenatal paternity test is mostly performed by using Amniocentesis or Chorionic Villus Sampling (CVS) methods. However, these methods require invasive procedures, which are potentially harmful for both the mother and the fetus. Currently, the invention of of Cell-Free Fetal DNA (cffDNA) has offered the opportunity of performing prenatal paternity test non-invasively. Materials and Methods : This study is a cross sectional descriptive study to detect cell free fetal DNA at 13 STR loci and at amelogenin gene to evaluate fetus gender, which will be compared to the baby gender afterbirth. Healthy third semester pregnant women were included as participants. Inform consent for both the mother and the biological father has been provided. Result : Four participants has been evaluated. In this study, in all participants, we found the presence of cffDNA in almost all of the STR loci. Some loci cannot be detected due to the small amount of cffDNA in the loci. All fetus genders detected by cffDNA in the amelogenin gene macthed the gender of the four babies afterbirth. Conclusion : The use of Cell-Free Fetal DNA (cffDNA) is a potential non-invasive methods in prenatal paternity test. Additionally, the ability of the method to evaluate fetus gender has been suggested.

Paternity Test Through Kinship Analysis as Forensic Identification Technique

Paternity tests is often faced with the unavailability of information from the father, mother, or child as a comparison in forensic DNA examination process. Therefore, comparisons with information from close family lines are needed, for example from the victim's siblings or the perpetrator if there are no comparisons from parents or siblings. This study was conducted by the Human Genetic Study Group of Airlangga University in its campus from January to April 2020. The aim of the study was to analyze the use of kinship analysis in paternity test through STR CODIS examination on siblings. This was an observational laboratory study with a temporary design. DNA sample extraction level and purity results were measured with the mean DNA sample level of 675±5.35ng/µL, while the purity values ranged from 1.05 to 1.86. The paternity test principle is based on comparison process between the parents’ alleles with the child’s alleles. However, if the parents’ alleles are not available, the siblings’ alleles can be used as a comparison for identification purpose, which is known as kinship analysis. Statistically, full siblings have a 2 alleles accuracy probability of [0.25] 25%, which was the same as not having the same allele or 0 allele, while 1 allele accuracy reached 50%. All CODIS STR loci had the highest percentage of 2 allele sharing. Therefore, it is recommended to use sibling or kinship analysis if both parents are absent.

Temporal variation in reproductive success and the effects of differential parental care on the progeny of fish with demersal eggs

A paternity test based on microsatellite analysis of eggs under the parental care of a bourgeois male Pacific sergeant major, Abudefduf troschelii, included gametes from five females and two parasitic males. The nest area was monitored for five days to evaluate variation in reproductive success, and an approximately 300% increase occurred on the first and second days after new oviposition followed by a more than 300% reduction from the second to fifth days due to predation or filial cannibalism. Finally, the egg area on the last day corresponded to the initial nest area, suggesting selective care by the male.

THE USE OF KINSHIP ANALYSIS ON PATERNITY TESTING THROUGH CODIS STR LOCI ‘CSF1PO’ AND ‘THO1’

Introduction: Paternity tests compare a child’s DNA pattern with the possible father to examine the DNA heritage in ensuring kinship. If there is no information from the father and mother or the child that can be used as a comparison in the forensic DNA examination process (paternity test), there must be a comparison from a close relative as an alternative to obtain the forensic DNA examination. This experiment’s purpose is to analyze the use of kinship analysis in forensic identification especially in a paternity test. Methods: This is a descriptive experiment using a cross-sectional design through locus analysis of DNA forensic examination in paternity test using the kinship analysis through STR CODIS loci: CSF1PO and THO1. The variables of this experiment were ‘locus’ as the independent variable and ‘allele number’ as the dependent variable. This experiment conducted a paternity tests to 8 samples in which the subjects were siblings; the test was administered on the CSF1PO and TH01 loci. Results: This experiment displayed similar allele numbers on the same locus, both in 50% and 100% allele numbers; the research showed similarity in allele numbers of both siblings of which ¼ were inherited from the parents (for 50% allele number similarity), and ½ were inherited from the parents (for 100% allele number similarity). Conclusion: This proves that a paternity test using siblings as the closest kin (kinship analysis) can be used as an alternative if no comparison is obtained from both the parents.

PELATIHAN PROSEDUR LABORATORIUM ANALISIS DNA

Currently, analysis based on DNA is one of the techniques for solving the problems such as paternity test, determining the identity of accident victims, natural disasters victims, and authentication of foods ingredients. But not everyone knows about the stages of DNA analysis laboratory procedures. The purpose of this workshop is to provide knowledgments about DNA analysis laboratory procedures. The training was attended by 30 participants from various universities and research centers in Riau Province. Workshop materials include DNA isolation of animal, plant and bacterial, electrophoresis, polymerase chain reaction (PCR), and visualization of DNA bands. Evaluation uses a questionnaire distributed to participants. Questionnaire analysis showed that workshop was very useful especially for beginners who want to be involved in DNA analysis activities in the laboratory.