A GEFI collaborative exercise on DNA/RNA co-analysis and mRNA profiling interpretation

Carnevali, E.; Lacerenza, Daniela; Severini, Simona; Alessandrini, Federica; Bini, Carla; Nunzio, C. Di; Nunzio, Mattia Di; Fabbri, M.; Fattorini, Paolo; Piccinini, A.; Ponzano, E.; Portera, G.; Previderé, Carlo; Renieri, Alessandra; Scarnicci, Francesca; Verzeletti, Andrea; Pelotti, Susi; Berge, M. van den; Sijen, Titia; Robino, Carlo

doi:10.1016/j.fsigss.2017.09.018

Cited by 9 publications

(11 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The function of CYP2B7P1 and MYOZ1 in vagina remains unknown and their expression in women from different age groups has not been previously investigated. The results obtained confirm that, in general, MYOZ1 is a less sensitive vaginal marker compared to MUC4 and CYP2B7P1, both in R and PM donors [3]. Overall, the three combined vaginal markers included in the 19-plex mRNA profiling assay appear highly efficient in identifying vaginal mucosa irrespective of the donors age category, even in non-recent samples (6-20 months).…”

Section: Discussionsupporting

confidence: 65%

“…Subsets of swabs (n = 13-14), designed as to minimize differences in the ratio between R and PM samples and mean time since swab collection (7.3 months ± 0.4 SD), were distributed to participating laboratories. RNA extraction, cDNA synthesis, mRNA profiling using a 19-plex mRNA assay [1] and scoring of results [2] was done according to protocols previously adopted in a preparatory GeFI collaborative exercise [3]. For capillary electrophoresis, each participant used the available instrumentation (310, 3130, 3500 and SeqStudio genetic analyzers), and the settings (analytical threshold, etc.)…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of vaginal mRNA markers in women from different age groups: A GeFI collaborative study

Robino

Chierto

Alessandrini³

et al. 2019

Forensic Science International: Genetics Supplement Series

Self Cite

View full text Add to dashboard Cite

Forensic mRNA profiling assays normally include a set of vaginal-specific markers. Although it is known that vagina undergoes characteristic age-related morphological and physiological changes over a lifetime, few studies have evaluated the efficacy of proposed forensic vaginal mRNA markers in women from different age groups. In this collaborative study involving ten GeFI (Italian working group of ISFG) laboratories, a 19-plex mRNA profiling assay including three vaginal-specific markers (CYP2B7P1, MUC4, MYOZ1) was tested in a collection of vaginal swabs obtained from female volunteer donors in their reproductive years (n = 84) and postmenopausa (n = 55). Differential expression of vaginal markers in the two age categories was assessed by means of: a) overall success rate of mRNA profiling (vaginal mucosa "observed" in the tested sample according to scoring protocol) b) average peak height ratios between vaginal-specific markers within mRNA profiling replicates. Other factors potentially influencing mRNA profiling outcomes, like time interval between vaginal swab collection and analysis, concurrence of menstrual cycle and recent sexual activity at the time of sampling were also investigated.

show abstract

Section: Discussionsupporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Evaluation of vaginal mRNA markers in women from different age groups: A GeFI collaborative study

Robino

Chierto

Alessandrini³

et al. 2019

Forensic Science International: Genetics Supplement Series

Self Cite

View full text Add to dashboard Cite

show abstract

“…Markers, primer sequences, and concentrations were adopted from Van den Berge et al [7]. This molecular system was previously tested in an ISFG Italian Working Group -GEFI collaborative work [8].All thermal cycling steps were accomplished in a Veriti® 96-Well Thermal Cycler (Thermo Scientific®).DNA isolate from the co-extraction process was typed using a conventional STR kit, represented by the AmpFlSTR® NGM PCR Amplification kit (Thermo Scientific®).Detection of all amplified fragments was performed using an ABI PRISM 310 Genetic Analyzer (Thermo Scientific®) and allele calling was accomplished using GeneMapper ID-X V1.0 software (Thermo Scientific®). Allele designation was carried out in comparison to control DNA 007 and allelic ladder provided by the manufacturer.…”

Section: Methodsmentioning

confidence: 99%

mRNA profiling in casework analyses

Fabbri¹,

Frisoni²,

Marini³

et al. 2020

JIOMICS

Self Cite

View full text Add to dashboard Cite

Our work involved the application of mRNA profiling to three bloodstains previously processed by the laboratory, in which routinely methods (immunochromatography) used for specific blood identification showed negative results. Analyses were accomplished on a sweater worn by the suspect. No bloodstains were found during the routine inspection. In order to identify possible latent bloodstains, luminol was applied. After the reagent vaporization, three distinct areas of luminescence appeared. In order to confirm the presence of the human hemoglobin, luminescent surfaces were collected using 4N6FLOQSwabs® and tested using HemDirect Hemoglobin test (SERATEC®). All samples analysed showed negative results. Due to the merging blood-specific markers mRNA profiling, the areas previously identified were sampled newly and tested using the three blood specific markers HBB, ALAS2 and CD93, together with two housekeeping genes represented by ACTB and 18S-rRNA. All samples showed positive results for all three blood specific mRNA markers.

show abstract

“…In recent decades, gene expression analysis of mRNA markers characteristic of body fluids has been used for the forensic identification of biological samples [ 8 , 9 , 10 , 11 , 12 , 13 ]. Those studies reported bactericidal permeability-increasing protein fold-containing family member A1 (BPIFA1) as a nasal secretion marker [ 14 , 15 ]. BPIFA1 is a lipid-binding protein that plays a role in the innate immune responses of the upper airway [ 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

A Proposed Procedure for Discriminating between Nasal Secretion and Saliva by RT-qPCR

Akutsu

Watanabe

2020

Diagnostics

View full text Add to dashboard Cite

In forensic casework, nasal secretion can be a good source of DNA. Moreover, saliva can prove useful in cases of sexual assault. However, discriminating between these body fluids is often difficult because of cross-reactivity between them on presumptive and confirmatory tests. Therefore, an RT-qPCR procedure was developed to discriminate between nasal secretion and saliva. Characteristic genes in nasal secretion and/or saliva (BPIFA1, STATH, HTN3, and PRH2) were selected as candidates. Discrimination criteria were established based on the expression levels of these markers in various body fluids. In addition, a flowchart was proposed and used to discriminate among nasal secretion, saliva, and other body fluids in various forensic samples. BPIFA1 was highly expressed in nasal secretion but was also expressed in saliva, semen, and vaginal fluid at trace levels. STATH was expressed in nasal secretion and saliva but not in other body fluids. HTN3 was specifically expressed in most of the saliva samples, as reported previously. Unexpectedly, PRH2 was expressed in only a few saliva samples. Using the proposed criteria and flowchart, nasal secretion and saliva were successfully discriminated among the various body fluids tested. The developed procedure could be useful in forensic casework.

show abstract

A GEFI collaborative exercise on DNA/RNA co-analysis and mRNA profiling interpretation

Cited by 9 publications

References 5 publications

Evaluation of vaginal mRNA markers in women from different age groups: A GeFI collaborative study

Evaluation of vaginal mRNA markers in women from different age groups: A GeFI collaborative study

mRNA profiling in casework analyses

A Proposed Procedure for Discriminating between Nasal Secretion and Saliva by RT-qPCR

Contact Info

Product

Resources

About