scholarly journals 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)

2021 ◽  
Vol 16 (1) ◽  
pp. 6-12
Author(s):  
Ahmad Yudianto ◽  
Arofi Kurniawan ◽  
Toetik Koesbardiati ◽  
Achmad Faisol ◽  
Fery Setiawan ◽  
...  

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.

2019 ◽  
Vol 19 (2) ◽  
pp. 105-111
Author(s):  
Nadia Shafei ◽  
Mohammad Saeed Hakhamaneshi ◽  
Massoud Houshmand ◽  
Siavash Gerayeshnejad ◽  
Fardin Fathi ◽  
...  

Background: Beta thalassemia is a common disorder with autosomal recessive inheritance. The most prenatal diagnostic methods are the invasive techniques that have the risk of miscarriage. Now the non-invasive methods will be gradually alternative for these invasive techniques. Objective: The aim of this study is to evaluate and compare the diagnostic value of two non-invasive diagnostic methods for fetal thalassemia using cell free fetal DNA (cff-DNA) and nucleated RBC (NRBC) in one sampling community. Methods: 10 ml of blood was taken in two k3EDTA tube from 32 pregnant women (mean of gestational age = 11 weeks), who themselves and their husbands had minor thalassemia. One tube was used to enrich NRBC and other was used for cff-DNA extraction. NRBCs were isolated by MACS method and immunohistochemistry; the genome of stained cells was amplified by multiple displacement amplification (MDA) procedure. These products were used as template in b-globin segments PCR. cff-DNA was extracted by THP method and 300 bp areas were recovered from the agarose gel as fetus DNA. These DNA were used as template in touch down PCR to amplify b-globin gen. The amplified b-globin segments were sequenced and the results compared with CVS resul. Results: The data showed that sensitivity and specificity of thalassemia diagnosis by NRBC were 100% and 92% respectively and sensitivity and specificity of thalassemia diagnosis by cff-DNA were 100% and 84% respectively. Conclusion: These methods with high sensitivity can be used as screening test but due to their lower specificity than CVS, they cannot be used as diagnostic test.


Author(s):  
Citra Manela ◽  

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.


2021 ◽  
Vol 6 (2) ◽  
pp. 60318
Author(s):  
Ahmad Yudianto ◽  
Fery Setiawan ◽  
Simon Martin Manyanza Nzilibili

Sibling indices can be used as a comparison through alleles Short Tandem Repeats [STR] loci. This is an observational study among Maduranese with 4 STR loci (CSF1PO, THOI, TPOX, vWA) obtained from their blood samples. The percentage of alleles shared: 82.5% [33 times] with 2 allele sharing, 12.5% [5 times] with 1 allele sharing, and 5 % [2 times] with 0 sharing alleles. Sibling indices (SI) calculation results: 65% of sibling indices pairs have SI greater than 100 and 15% of them were between 10-100 (strong and very strong). Sibling indices interpretation is supported; therefore, the claimed sibling indices relationships were indeed true among Maduranese ethnic group in Surabaya.


2019 ◽  
Vol 2 (2) ◽  
pp. 113
Author(s):  
Dwi Fitrianti Arieza Putri ◽  
Ahmad Yudianto

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.


2021 ◽  
Vol 53 (1) ◽  
Author(s):  
Ahmad Yudianto ◽  
Fery Setiawan ◽  
Reni Sumino

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.


2020 ◽  
Vol 16 (2) ◽  
pp. 138-152
Author(s):  
Bingren Zhang ◽  
Chu Wang ◽  
Chanchan Shen ◽  
Wei Wang

Background: Responses to external emotional-stimuli or their transitions might help to elucidate the scientific background and assist the clinical management of psychiatric problems, but pure emotional-materials and their utilization at different levels of neurophysiological processing are few. Objective: We aimed to describe the responses at central and peripheral levels in healthy volunteers and psychiatric patients when facing external emotions and their transitions. Methods: Using pictures and sounds with pure emotions of Disgust, Erotica, Fear, Happiness, Neutral, and Sadness or their transitions as stimuli, we have developed a series of non-invasive techniques, i.e., the event-related potentials, functional magnetic resonance imaging, excitatory and inhibitory brainstem reflexes, and polygraph, to assess different levels of neurophysiological responses in different populations. Results: Sample outcomes on various conditions were specific and distinguishable at cortical to peripheral levels in bipolar I and II disorder patients compared to healthy volunteers. Conclusions: Methodologically, designs with these pure emotions and their transitions are applicable, and results per se are specifically interpretable in patients with emotion-related problems.


2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Andy Sombke ◽  
Carsten H. G. Müller

Abstract Background The jointed appendage is a key novelty in arthropod evolution and arthropod legs are known to vary enormously in relation to function. Among centipedes, the ultimate legs always are distinctly different from locomotory legs, and different centipede taxa evolved different structural and functional modifications. In Geophilomorpha (soil centipedes), ultimate legs do not participate in locomotion and were interpret to serve a sensory function. They can be sexually dimorphic and in some species, male ultimate legs notably appear “hairy”. It can be assumed that the high abundance of sensilla indicates a pronounced sensory function. This study seeks for assessing the sensory diversity, however, documents the surprising and unique case of an extensive glandular epithelium in the ultimate legs of three phylogenetically distant species. Results The tightly aggregated epidermal glands with stalked ducts – mistakenly thought to be sensilla – were scrutinized using a multimodal microscopic approach comprising histology as well as scanning and transmission electron microscopy in Haplophilus subterraneus. Hence, this is the first detailed account on centipede ultimate legs demonstrating an evolutionary transformation into a “secretory leg”. Additionally, we investigated sensory structures as well as anatomical features using microCT analysis. Contrary to its nomination as a tarsus, tarsus 1 possesses intrinsic musculature, which is an indication that this podomere might be a derivate of the tibia. Discussion The presence and identity of ultimate leg associated epidermal glands with stalked ducts is a new discovery for myriapods. A pronounced secretory as well as moderate sensory function in Haplophilus subterraneus can be concluded. The set of characters will improve future taxonomic studies, to test the hypotheses whether the presence of these specialized glands is a common feature in Geophilomorpha, and that tarsus 1 may be a derivate of the tibia. As the number of epidermal glands with stalked ducts is sexually dimorphic, their function might be connected to reproduction or a sex-specific defensive role. Our results, in particular the unexpected discovery of ‘glandular hairs’, may account for a striking example for how deceptive morphological descriptions of epidermal organs may be, if based on non-invasive techniques alone.


Biosensors ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 189
Author(s):  
David Bamgboje ◽  
Iasonas Christoulakis ◽  
Ioannis Smanis ◽  
Gaurav Chavan ◽  
Rinkal Shah ◽  
...  

Diabetes mellitus (DM) is a chronic disease that must be carefully managed to prevent serious complications such as cardiovascular disease, retinopathy, nephropathy and neuropathy. Self-monitoring of blood glucose is a crucial tool for managing diabetes and, at present, all relevant procedures are invasive while they only provide periodic measurements. The pain and measurement intermittency associated with invasive techniques resulted in the exploration of painless, continuous, and non-invasive techniques of glucose measurement that would facilitate intensive management. The focus of this review paper is the existing solutions for continuous non-invasive glucose monitoring via contact lenses (CLs) and to carry out a detailed, qualitative, and comparative analysis to inform prospective researchers on viable pathways. Direct glucose monitoring via CLs is contingent on the detection of biomarkers present in the lacrimal fluid. In this review, emphasis is given on two types of sensors: a graphene-AgNW hybrid sensor and an amperometric sensor. Both sensors can detect the presence of glucose in the lacrimal fluid by using the enzyme, glucose oxidase. Additionally, this review covers fabrication procedures for CL biosensors. Ever since Google published the first glucose monitoring embedded system on a CL, CL biosensors have been considered state-of-the-art in the medical device research and development industry. The CL not only has to have a sensory system, it must also have an embedded integrated circuit (IC) for readout and wireless communication. Moreover, to retain mobility and ease of use of the CLs used for continuous glucose monitoring, the power supply to the solid-state IC on such CLs must be wireless. Currently, there are four methods of powering CLs: utilizing solar energy, via a biofuel cell, or by inductive or radiofrequency (RF) power. Although, there are many limitations associated with each method, the limitations common to all, are safety restrictions and CL size limitations. Bearing this in mind, RF power has received most of the attention in reported literature, whereas solar power has received the least attention in the literature. CLs seem a very promising target for cutting edge biotechnological applications of diagnostic, prognostic and therapeutic relevance.


2020 ◽  
Vol 8 (1) ◽  
Author(s):  
Klara Retko ◽  
Maša Kavčič ◽  
Lea Legan ◽  
Polonca Ropret ◽  
Bojana Rogelj Škafar ◽  
...  

AbstractIn this study, a painted beehive panel from the collection of the Slovene Ethnographic Museum was examined with respect to its material composition with the aim to reveal the painting technique. Due to the state of degradation due to outdoor weathering (UV irradiation, rainfall, extreme temperature and humidity fluctuations), as well as past conservation interventions, the object represented a complex analytical challenge. We aimed for non-invasive techniques (FTIR in reflection mode, Raman spectroscopy and hyperspectral imaging in the range of 400–2500 nm); however, in order to explore paint layers, cross-sections were also analysed using Raman spectroscopy. FTIR spectroscopy in transmission mode and gas chromatography coupled to mass spectrometry were also used on sample fragments. Various original materials were identified such as pigments and binders. The surface coating applied during conservation interventions was also characterised. Additionally, organic compounds were found (oxalate, carboxylate), representing transformation products. The potential use of Prussian blue as a background paint layer is discussed.


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