New cyt b gene universal primer set for forensic analysis

López-Oceja, A.; Gamarra, David; Borragan, S.; Jiménez-Moreno, Susana; Pancorbo, Marian M. de

doi:10.1016/j.fsigen.2016.05.001

Cited by 37 publications

(21 citation statements)

References 34 publications

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“…To verify that the origin of endothelial cells in the long-term experimental group, we extracted the endothelial cell DNA for PCR and sequencing. Nucleotide sequences of human and animal mitochondrial cytochrome b genes are known to be species-speci c and have been used for species identi cation in forensic investigations [22,23]. In order to distinguish human and monkey species, we designed speci c Cytb primers.…”

Section: Discussionmentioning

confidence: 99%

Long-term observation and sequencing analysis of SKPs-derived corneal endothelial cell-like cells for treating corneal endothelial dysfunction

Shen

Sun

et al. 2020

Preprint

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Background Corneal endothelial dysfunction is a principal cause of visual deficiency. Corneal transplantation is the most effective treatment for corneal endothelial dysfunction. However, a severe shortage of available donor corneas remains a global challenge. Previously, we acquired corneal endothelial cell-like cells (CEC-like cells) derived from skin-derived precursors (SKPs). CEC-like cells exerted excellent therapeutic effect in rabbit and monkey corneal endothelial dysfunction models. Method: We prolonged the observation in the monkey experiment for 2 years. Further transcriptome sequencing was also carried out on HCEC, CEC-like cells before transplantation and after transplantation. Results We confirmed that the monkey cornea remained transparent and the CEC-like cells could adapt to the microenvironment after transplantation. The gene expression pattern of CEC-like cells was similar to HCEC and changed slightly after transplantation. Conclusions This study presented a brand-new insight into CEC-like cells and further provided a promising prospect of cell-based therapy for corneal endothelial dysfunction. The renewable cell source, novel derivation method and simple treatment strategy may be clinically applied in regenerative medicine in the future.

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Section: Discussionmentioning

confidence: 99%

Long-term observation and sequencing analysis of SKPs-derived corneal endothelial cell-like cells for treating corneal endothelial dysfunction

Shen

Sun

et al. 2020

Preprint

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“…From the consensus sequences, we performed an alignment of the consensus sequences using Mafft (using the L-INS-i algorithm). We selected a region with high differentiation between species guided by the species-variable regions identified in (47), and then entered that region into the Integrated DNA Technologies, Inc. (IDT, Coralville, Iowa) PrimerQuest online tool along with ddPCR conditions specified by Bio-Rad to design forward primer, reverse primer, and probe sequences.…”

Section: Methodsmentioning

confidence: 99%

A Novel Droplet Digital PCR Human mtDNA Assay for Global Fecal Source Tracking

Zhu

Suttner

Konstantinidis

et al. 2019

Preprint

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229 / 250 words max.) 6 Human mitochondrial DNA (mtDNA) provides a promising target for microbial source tracking 7 because it is unique to humans and universal across human individuals. We developed a droplet 8 digital PCR (ddPCR) assay, hCYTB484, targeting the cytochrome b gene of the human mtDNA 9 and compared the performance of the hCYTB484 assay with a widely used assay targeting 10 human-associated Bacteroides, the HF183/BacR287 assay. We also defined and validated the 11 analytical limit of detection and analytical lower limit of quantification; these analytical limits 12 determine the concentration levels above which samples are declared to be positive and 13 quantifiable for the target, respectively. We found both assays to be highly specific (95%) 14 against cow and pig feces; however, the hCYTB484 was more sensitive when tested against 15 individual human feces from US, Bangladesh, and Mozambique (100% versus a mean of 51% 16 across the 3 countries). To further compare the performance of the two assays, we utilized a 17 receiver operating characteristic curve, showing that the hCYTB484 marker was widely 18 distributed across human feces from the 3 different geographical regions tested and in higher 19 quantities than the HF183/BacR287 marker. The higher performance of the hCYTB484 marker 20 in individual feces is a desirable characteristic in the detection of fecal pollution from sources to 21 which fewer individuals contribute, such as non-sewered types of sanitation that serve most of 22 Earth's population and carry the highest risk of exposure to fecal-oral pathogens. 23 Importance (142/150 words) 24 The usefulness of an ideal human-specific, culture-and library-independent marker to the 25 microbial source tracking field is reflected by the numerous efforts to develop such markers; 26 however, thus far, microbial-based markers of this type have exhibited variable source-27 specificity across geographies and variable sensitivity across scales of fecal waste. Most of the 28 3 world's population is served by non-sewered forms of sanitation that also carry the highest risk 29 of exposure to fecal-oral pathogens. This reality underscores the need for markers of human fecal 30 contamination that have high sensitivity in fecal pollution sources to which fewer individuals 31 contribute to, such as fecal sludges found in pit latrines. We show that human mtDNA-based 32 methods can be highly source-specific and highly sensitive in smaller scales of fecal pollution, 33 providing a useful addition to the microbial source tracking toolbox by complementing the 34 variable performance of microbial-based markers. 35 4 Word Count 3854/6000 (should not exceed approximately 6,000 words, exclusive of methods, 36 references, figure legends, tables, and supplemental material) 37 Introduction 38Within the microbial source tracking (MST) field, the development of an ideal human-specific, 39 culture-and library-independent marker has been a goal for many research efforts for a variety of 40 reasons-nota...

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“…Cytochrome b markers have shown good feasibility in detecting species of origin from the sample mixtures. The nucleotide sequence of the cytochrome b gene contains species-specific information and has been used in food safety as well as in forensic investigations in a number of studies [3,4,5,6].…”

Section: Introductionmentioning

confidence: 99%

Identification Human and Animal Blood Mixtures Using Human Cytochrome b Gene

Widodo¹,

Furqoni²,

Yudianto³

et al. 2018

Proceedings of the 1st International Conference Postgraduate School Universitas Airlangga : "Implementation of Climate Change A

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Abstract-Human and animal blood has a similar composition so it is difficult to distinguish between the two. The aim of this study was to identify human blood from human and animal blood mixtures using the cytochrome b gene that has a species-specific DNA sequence. The DNA extracted from varied human and chicken blood was amplified using a human-specific cytochrome b gene primer. The result showed that 10 % human blood from the mixtures could be identified and the DNA extracted from the human blood could be amplified. The concentration of 0.01 ng still showed the appropriate DNA band. The primer was very sensitive and specific, so it useful for forensic purposes to verify human blood in a blood mixture.

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New cyt b gene universal primer set for forensic analysis

Cited by 37 publications

References 34 publications

Long-term observation and sequencing analysis of SKPs-derived corneal endothelial cell-like cells for treating corneal endothelial dysfunction

Long-term observation and sequencing analysis of SKPs-derived corneal endothelial cell-like cells for treating corneal endothelial dysfunction

A Novel Droplet Digital PCR Human mtDNA Assay for Global Fecal Source Tracking

Identification Human and Animal Blood Mixtures Using Human Cytochrome b Gene

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