scholarly journals Comparison of Time Course Detection of Human Male DNA from Blood Stains on Various Objects on Surface in a Natural Environment and in a Laboratory Using Loop-Mediated Isothermal Amplification (LAMP)

Scientifica ◽  
2021 ◽  
Vol 2021 ◽  
pp. 1-7
Author(s):  
Ratchanok Kumsiri ◽  
Panan Kanchanaphum

In forensic study, the biological evidence can easily degrade, especially DNA. Degraded and environmentally challenged samples can produce numerous problems in forensic DNA analysis including loss of band product. Loop-mediated isothermal amplification or LAMP is one of the DNA analysis techniques used in forensic study. This study explores the limitations of the efficiency of the LAMP technique on abandoned DNA. For the DNA template, 8 male and 2 female blood-stained samples were taken from the surfaces, namely, brick, cloth, and tile from inside, and buried outside the laboratory. The LAMP reaction was used to amplify the SRY gene for detecting male DNA. All the blood-stained samples were stored for 1, 7, 15, 30, and 45 day (s). The LAMP product from the blood-stained samples on all the surfaces that were kept in a laboratory was detected using the gel electrophoresis technique from day 1 until day 45. However, the LAMP product on day 30 and 45 was smear and dim. The LAMP product from the blood-stained samples buried outside the laboratory was observed using the gel electrophoresis technique within day 30 (smear and dim). To increase the efficiency of detection, the qLAMP technique detected product on all the male samples from all the surfaces buried outside the laboratory for 45 days. The results indicate that this LAMP condition was possible detecting male DNA and the environmental factors are the main influence on the sensitivity of the LAMP technique. In addition, the qLAMP technique can increase the capacity and sensitivity of the detection.

2018 ◽  
Vol 2018 ◽  
pp. 1-8
Author(s):  
Panan Kanchanaphum

This study explores determining the sex of humans from blood stains taken from different surfaces and compares the time course of detection with the conventional PCR, Conventional Loop Mediated Isothermal Amplification (LAMP), and LAMP-Lateral Flow Dipstick (LFD). For the DNA templates, 7 male and 7 female blood stained samples were extracted and added to LAMP and PCR reaction solution to amplify the SRY gene. The DNA samples were extracted from the following blood stained materials: cloth, wood, clay, and tile. Then, the samples were stored at room temperature for 1, 7, 30, and 60 day(s). After the DNA amplification, the gel electrophoresis process was applied to detect LAMP product. The LFD was combined with the LAMP to detect LAMP product on the male cloth samples. For the male samples, the time course of detection on the first and seventh days indicated positive for both LAMP and PCR products on all the surfaces while no DNA amplification was found on any of the female samples. On day 30, positive LAMP product was still found on all the male samples. However, it had faded on the tiles. Moreover, all the male samples, which had tested positive for PCR product, were blurred and unclear. On day 60, LAMP product was still found on all the male samples. Conversely, the PCR method resulted in no bands showing for any of the male samples. However, the LAMP-LFD method detected product on all the male samples of cloth. The results show that the LAMP is an effective, practical, and reliable molecular-biological method. Moreover, the LFD can increase the efficiency and sensitivity of the LAMP, making it more suitable for field studies because gel electrophoresis apparatus is not required.


2021 ◽  
Author(s):  
Pankaj Shrivastava ◽  
R. K. Kumawat ◽  
Pushpesh Kushwaha ◽  
Akshay Kumar ◽  
Manisha Rana ◽  
...  

Abstract Ever since the inception of STR-based forensic DNA analysis for human Identification and DNA profiling, the approach developed continuously. Most of these developments are pertaining to achieve the best polymorphic loci set for analysis, high inhibitor tolerance capability, increase in sensitivity, reduction in turnaround time, and cost-effectiveness. On the similar line, the SF25TM PCR amplification kit, a 6 fluorescent dye-chemistry based autosomal STR kit was developed, which analyses 25 STR loci viz., FAM fluorescent dye-labelled D3S1358, D13S317, D7S820, D16S539, D1S1656, and Penta E; HEX fluorescent dye-labelled TPOX, TH01, D2S1338, CSF1PO, and Penta D; SUM fluorescent dye-labeled D19S433, vWA, D21S11, D18S51, and D6S1043; LYN fluorescent dye-labelled Amelogenin, D8S1179, D5S818, D12S391, and FGA; PUR fluorescent dye-labelled D22S1045, Y indel; D2S441 and D10S1248, The developmental validation of the SF25 PCR Amplification reagents were carried out in this study to analyze its specificity, sensitivity, and concordance. This multiplex kit showed higher sensitivity with 125pg of DNA template generating a complete profile (with 100% allele call). Out of 83 samples, 10 samples showed aberrant triallelic pattern/aberrant heterozygous pattern at D16S539 locus. The possible discrepant homozygous condition at locus D1S1656 might be due to the invasion of alleles at D1S1656 locus present adjacent to D16S539 locus. Such invasion resulted due to the shortening of the allelic bin at the D1S1656 locus. The allelic ladder of the SF25TM PCR amplification kit reveals the allelic range of D1S1656 from 11 to 19.3 which can be interpreted according to the casework as deemed by the end-users for their samples.


2019 ◽  
Vol 39 (2) ◽  
Author(s):  
Qing Huang ◽  
Zhiwu Li ◽  
Zhiguo Ma ◽  
He Li ◽  
Runqian Mao

Abstract Guang-dilong (Pheretima aspergillum) is a traditional Chinese animal medicine that has been used for thousands of years in China. In the present study, we purposed to establish a new rapid identification method for Guang-dilong. We provided a useful technique, loop-mediated isothermal amplification (LAMP), to differentiate Guang-dilong from other species. Four specific LAMP primers were designed based on mitochondrial cytochrome c oxidase I (COI) gene sequences of Guang-dilong. LAMP reaction, containing DNA template, four primers, 10× Bst DNA polymerase reaction buffer, dNTPs, MgSO4, and Bst DNA polymerase, was completed within 60 min at 63°C. The LAMP product can be visualized by adding SYBR Green I or detected by 2% gel electrophoresis. LAMP technology was successfully established for rapid identification of Guang-dilong. In addition, DNA template concentration of 675 fg/μl was the detection limit of LAMP in Guang-dilong, which was 1000-times higher than conventional PCR. The simple, sensitive, and convenient LAMP technique is really suited for on-site identification of Guang-dilong in herbal markets.


2015 ◽  
Vol 15 ◽  
pp. 98-104 ◽  
Author(s):  
Sergey Borovko ◽  
Alena Shyla ◽  
Victorya Korban ◽  
Alexandra Borovko

2018 ◽  
Vol 59 (1) ◽  
pp. 9-16 ◽  
Author(s):  
Samuele Manzoni ◽  
Andrea Ossoli ◽  
Venusia Cortellini ◽  
Andrea Verzeletti

Forensic examination of human remains is a complex process that relies on the contribution of multidisciplinary forensic medicine specialties. Here we present a complex forensic case regarding a double murder whose victims were found almost completely skeletonized. Post-mortem investigations allowed us to define the biological profile of the two bodies (ancestry, sex, age and stature), to discover their identity through forensic DNA analysis, and to detect peri-mortem injuries caused by firearms and stabbing weapons. Three men were recognized as involved in the crime and two of them were condemned to life imprisonment for homicide. The judges accepted the reconstruction of the crime promoted by the Prosecutor (double firearm murder).


BioTechniques ◽  
2009 ◽  
Vol 47 (5) ◽  
pp. 951-958 ◽  
Author(s):  
Johannes Hedman ◽  
Anders Nordgaard ◽  
Birgitta Rasmusson ◽  
Ricky Ansell ◽  
Peter Rådström

2020 ◽  
Vol 77 (2) ◽  
pp. 93-99
Author(s):  
Р. Л. Степанюк ◽  
С. І. Перлін

The authors of the article have studied the problems and perspectives of the formation of specific branch of forensic technology, which is devoted to DNA analysis in order to solve the tasks arising in criminal proceeding. Particular attention has been paid to the lack of a corresponding component in the domestic system of forensic technology, unlike the forensic science of foreign countries. The necessity of development of forensic DNA analysis as an independent branch of forensic technology has been argued. It is confirmed by the following main arguments: the methodology of this field of research is based on the theory of forensic identification; its objects are traces of human and other living organisms; DNA analysis technologies are developed using the achievements of different sciences and adapted to solve problems of crime detection and investigation; they are aimed to ensuring the activities of law enforcement agencies in counteracting crime; the scope of DNA analysis application in crime combating should not be limited to forensic activities; legislative regulation of collecting and using personal genetic data is essential; DNA analysis technologies in terms of practical significance and fundamental scientific basis exceed all other branches of forensic technology. The authors have offered to define forensic DNA analysis as the branch of forensic technology that studies individual genetic characteristics of living organisms contained in their DNA, in order to identify them and solve diagnostic tasks in the detection and investigation of criminal offenses. Its structure has been determined. The authors have provided perspective development directions of forensic DNA analysis: ensuring the appropriate state of legislative regulation of relations in the field of selection and use of personal genetic information; implementation of effective means and methods of detection and removal of biological traces and samples; improvement of methods of forensic DNA testing; formation of recommendations concerning the peculiarities of using DNA analysis results for proving; development of the latest technologies of forensic DNA analysis.


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