Detection of Saliva by Immunoaffinity and Mass Spectrometry

NCT ID: NCT06183385

Last Updated: 2023-12-27

Study Results

Results pending

The study team has not published outcome measurements, participant flow, or safety data for this trial yet. Check back later for updates.

Basic Information

Get a concise snapshot of the trial, including recruitment status, study phase, enrollment targets, and key timeline milestones.

Recruitment Status

NOT_YET_RECRUITING

Clinical Phase

NA

Total Enrollment

35 participants

Study Classification

INTERVENTIONAL

Study Start Date

2024-01-15

Study Completion Date

2024-12-31

Brief Summary

Review the sponsor-provided synopsis that highlights what the study is about and why it is being conducted.

The identification of saliva in genital area during a criminal investigation can be a critical component in the prosecution of a sexual assault in France, as non-consensual oral-genital intercourses have been considered as crimes since 2021.The development of highly specific methods for saliva detection is therefore crucial as the commonly employed screening methods lack specificity. Protein mass spectrometry has proven to be a sensitive and specific method but is particularly time consuming. A faster and more sensitive hybrid approach using automated immunoaffinity mass spectrometry (IP-LC-MS/MS) has been recently developed and has been found to be particularly performant for the detection of a seminal fluid protein (semenogelin), allowing a high-throughput seminal fluid identification in semen samples. Like semenogelin, specific salivary proteins such as histatin type 1, cystatin D or proline-rich proteins (PRPs) could be detected using this promising approach, which has never been tested on saliva samples. In collaboration with the Clinical Proteomics Platform and the Department of Reproductive Medicine of the University Hospital of Montpellier, we aim to develop a protocol for the detection of specific saliva proteins by IP-LC-MS/MS in sexual assault-type samples.

Detailed Description

Dive into the extended narrative that explains the scientific background, objectives, and procedures in greater depth.

Each participant will be contacted by a phone call the day before the visit, to present for a presentation of the study.The day of the visit, two types of samples will be collected :

* Saliva samples: on healthy volunteers in the Department of Legal Medicine (2 samples of 1.5 - 2 mL are collected for each volunteer).
* Vaginal samples : on women consulting in the Department of Reproductive Medicine (2 dry swabs on each patient). 2 groups, One group with vaginal secretion fluid samples only, and one group with vaginal secretion fluid + sperm samples.

Conditions

See the medical conditions and disease areas that this research is targeting or investigating.

Sexual Assault

Keywords

Explore important study keywords that can help with search, categorization, and topic discovery.

saliva proteins biomarkers mass spectrometry

Study Design

Understand how the trial is structured, including allocation methods, masking strategies, primary purpose, and other design elements.

Allocation Method

NON_RANDOMIZED

Intervention Model

PARALLEL

Primary Study Purpose

OTHER

Blinding Strategy

NONE

Study Groups

Review each arm or cohort in the study, along with the interventions and objectives associated with them.

Saliva

Collection of two saliva samples (approx. 1.5 to 2 mL each) by passive drooling in healthy volunteers

These samples will be centrifuged and the supernatant collected.

Group Type OTHER

Saliva collection

Intervention Type OTHER

Collection of 2 samples of 1.5 to 2mL of saliva by passive salivation in healthy volunteers

IP-LC-MS/MS : immunoprecipitation enrichment with liquid chromatography coupled to tandem mass spectrometry

Intervention Type OTHER

sample preparation and analysis :

* Impregnation of the tips (absorbent cotton) of sterile dry swabs and vaginal swabs with controlled quantities of saliva.
* Immunocapture of salivary proteins of interest (histatin type 1, cystatin D, PRPs) by protein A affinity purification
* LC-MS/MS analysis of eluted purified peptides (Multiple Reaction Monitoring mode)

Dilutions will be made from 10 to 10 (1/10, 1/100, 1/1000...), then after reaching a detectability threshold, specified by a second more precise analysis (e.g. if no signal at 1/1000, analysis at 1/500 then 1/250 etc.). Analytical sensitivity will be tested on three samples of each type (salivary, vaginal, vaginal + semen) to ensure reproducibility of results.

Operators, unaware of the presence of saliva in the samples, will then carry out saliva-specific protein detection analysis using the IP-LC-MS/MS method.

Vaginal secretions

In order to be as close as possible to real life, two vaginal swabs (introduced through the vaginal orifice over a length of 2 to 3 cm) will be taken with a sterile dry swab during the gynecological examination (prior to endovaginal ultrasound or any other endovaginal procedure) of patients included in the Reproductive Medicine Service.

Group Type OTHER

Vaginal secretion collection

Intervention Type OTHER

Collection of vaginal secretions with 2 dry swabs in women

IP-LC-MS/MS : immunoprecipitation enrichment with liquid chromatography coupled to tandem mass spectrometry

Intervention Type OTHER

sample preparation and analysis :

* Impregnation of the tips (absorbent cotton) of sterile dry swabs and vaginal swabs with controlled quantities of saliva.
* Immunocapture of salivary proteins of interest (histatin type 1, cystatin D, PRPs) by protein A affinity purification
* LC-MS/MS analysis of eluted purified peptides (Multiple Reaction Monitoring mode)

Dilutions will be made from 10 to 10 (1/10, 1/100, 1/1000...), then after reaching a detectability threshold, specified by a second more precise analysis (e.g. if no signal at 1/1000, analysis at 1/500 then 1/250 etc.). Analytical sensitivity will be tested on three samples of each type (salivary, vaginal, vaginal + semen) to ensure reproducibility of results.

Operators, unaware of the presence of saliva in the samples, will then carry out saliva-specific protein detection analysis using the IP-LC-MS/MS method.

Interventions

Learn about the drugs, procedures, or behavioral strategies being tested and how they are applied within this trial.

Saliva collection

Collection of 2 samples of 1.5 to 2mL of saliva by passive salivation in healthy volunteers

Intervention Type OTHER

Vaginal secretion collection

Collection of vaginal secretions with 2 dry swabs in women

Intervention Type OTHER

IP-LC-MS/MS : immunoprecipitation enrichment with liquid chromatography coupled to tandem mass spectrometry

sample preparation and analysis :

* Impregnation of the tips (absorbent cotton) of sterile dry swabs and vaginal swabs with controlled quantities of saliva.
* Immunocapture of salivary proteins of interest (histatin type 1, cystatin D, PRPs) by protein A affinity purification
* LC-MS/MS analysis of eluted purified peptides (Multiple Reaction Monitoring mode)

Dilutions will be made from 10 to 10 (1/10, 1/100, 1/1000...), then after reaching a detectability threshold, specified by a second more precise analysis (e.g. if no signal at 1/1000, analysis at 1/500 then 1/250 etc.). Analytical sensitivity will be tested on three samples of each type (salivary, vaginal, vaginal + semen) to ensure reproducibility of results.

Operators, unaware of the presence of saliva in the samples, will then carry out saliva-specific protein detection analysis using the IP-LC-MS/MS method.

Intervention Type OTHER

Eligibility Criteria

Check the participation requirements, including inclusion and exclusion rules, age limits, and whether healthy volunteers are accepted.

Inclusion Criteria

Saliva Samples

* Men and women aged 18 or more

Vaginal samples :

* Women aged 18 or more
* No unprotected vaginal sexual intercourse during the 10 days prior to the visit (group 1)
* Unprotected vaginal sexual intercourse with ejaculation in the 24 hours prior to the visit (group 2)

Exclusion Criteria

Saliva Samples :

* Active pathology of the saliva glands (infection, tumor)
* Unprotected oral-genital sexual intercourse in the 24 hours prior to the visit
* Failure to obtain written informed consent after a reflection period
* Pregnant or breast feeding women
* Adult protected by law or under guardianship or curatorship
* No affiliation to the French Social Security System or no benefit from such a system

Vaginal Samples :

* Oral-genital sexual intercourse (cunnilingus) in the 24 hours prior to the visit
* Failure to obtain written informed consent after a reflection period
* Pregnant or Breastfeeding women
* Adult protected by law or under guardianship or curatorship
* No affiliation to the French Social Security System or no benefit from such a system
Minimum Eligible Age

18 Years

Eligible Sex

ALL

Accepts Healthy Volunteers

Yes

Sponsors

Meet the organizations funding or collaborating on the study and learn about their roles.

University Hospital, Montpellier

OTHER

Sponsor Role lead

Responsible Party

Identify the individual or organization who holds primary responsibility for the study information submitted to regulators.

Responsibility Role SPONSOR

Principal Investigators

Learn about the lead researchers overseeing the trial and their institutional affiliations.

Pierre-Antoine PEYRON, PI

Role: STUDY_DIRECTOR

University Hospital, Montpellier

Locations

Explore where the study is taking place and check the recruitment status at each participating site.

PEYRON

Montpellier, , France

Site Status

Countries

Review the countries where the study has at least one active or historical site.

France

Central Contacts

Reach out to these primary contacts for questions about participation or study logistics.

Pierre-Antoine PEYRON, PI

Role: CONTACT

Phone: 04 67 33 85 86

Email: [email protected]

Laëtitia LEVEQUE

Role: CONTACT

Phone: 04 67 33 85 86

Email: [email protected]

References

Explore related publications, articles, or registry entries linked to this study.

LOI n°2021-478 du 21 avril 2021 visant à protéger les mineurs des crimes et délits sexuels et de l'inceste. JORF n°0095 du 22 avril 2021. Texte n°4

Reference Type BACKGROUND

Wornes DJ, Speers SJ, Murakami JA. The evaluation and validation of Phadebas(R) paper as a presumptive screening tool for saliva on forensic exhibits. Forensic Sci Int. 2018 Jul;288:81-88. doi: 10.1016/j.forsciint.2018.03.049. Epub 2018 Apr 20.

Reference Type BACKGROUND
PMID: 29734053 (View on PubMed)

Vandenberg N, van Oorschot RA. The use of Polilight in the detection of seminal fluid, saliva, and bloodstains and comparison with conventional chemical-based screening tests. J Forensic Sci. 2006 Mar;51(2):361-70. doi: 10.1111/j.1556-4029.2006.00065.x.

Reference Type BACKGROUND
PMID: 16566772 (View on PubMed)

Blanchard Muller M., Baccino E., Peyron P.A. Détection de traces salivaires dans le cadre d'agressions sexuelles : intérêt de l'utilisation d'une méthode immuno-chromatographique rapide (RSID-SalivaTest®) selon un protocole simplifié. Rev Médecine Légale 2023.

Reference Type BACKGROUND

Davidovics R, Saw YL, Brown CO, Prinz M, McKiernan HE, Danielson PB, Legg KM. High-throughput seminal fluid identification by automated immunoaffinity mass spectrometry. J Forensic Sci. 2022 May;67(3):1184-1190. doi: 10.1111/1556-4029.14975. Epub 2022 Jan 13.

Reference Type BACKGROUND
PMID: 35023573 (View on PubMed)

Sato H, Nakajima D, Ishikawa M, Konno R, Nakamura R, Ohara O, Kawashima Y. Evaluation of the Suitability of Dried Saliva Spots for In-Depth Proteome Analyses for Clinical Applications. J Proteome Res. 2022 May 6;21(5):1340-1348. doi: 10.1021/acs.jproteome.2c00099. Epub 2022 Apr 21.

Reference Type BACKGROUND
PMID: 35446574 (View on PubMed)

Yang H, Zhou B, Deng H, Prinz M, Siegel D. Body fluid identification by mass spectrometry. Int J Legal Med. 2013 Nov;127(6):1065-77. doi: 10.1007/s00414-013-0848-1. Epub 2013 Mar 24.

Reference Type BACKGROUND
PMID: 23525663 (View on PubMed)

Brown CO, Robbins BL, McKiernan HE, Danielson PB, Legg KM. Direct seminal fluid identification by protease-free high-resolution mass spectrometry. J Forensic Sci. 2021 May;66(3):1017-1023. doi: 10.1111/1556-4029.14646. Epub 2020 Dec 8.

Reference Type BACKGROUND
PMID: 33289932 (View on PubMed)

Legg KM, Powell R, Reisdorph N, Reisdorph R, Danielson PB. Discovery of highly specific protein markers for the identification of biological stains. Electrophoresis. 2014 Nov;35(21-22):3069-78. doi: 10.1002/elps.201400125. Epub 2014 Oct 1.

Reference Type BACKGROUND
PMID: 25070728 (View on PubMed)

Van Steendam K, De Ceuleneer M, Dhaenens M, Van Hoofstat D, Deforce D. Mass spectrometry-based proteomics as a tool to identify biological matrices in forensic science. Int J Legal Med. 2013 Mar;127(2):287-98. doi: 10.1007/s00414-012-0747-x. Epub 2012 Jul 29.

Reference Type BACKGROUND
PMID: 22843116 (View on PubMed)

Illiano A, Arpino V, Pinto G, Berti A, Verdoliva V, Peluso G, Pucci P, Amoresano A. Multiple Reaction Monitoring Tandem Mass Spectrometry Approach for the Identification of Biological Fluids at Crime Scene Investigations. Anal Chem. 2018 May 1;90(9):5627-5636. doi: 10.1021/acs.analchem.7b04742. Epub 2018 Apr 13.

Reference Type BACKGROUND
PMID: 29579379 (View on PubMed)

Legg KM, Powell R, Reisdorph N, Reisdorph R, Danielson PB. Verification of protein biomarker specificity for the identification of biological stains by quadrupole time-of-flight mass spectrometry. Electrophoresis. 2017 Mar;38(6):833-845. doi: 10.1002/elps.201600352. Epub 2017 Jan 30.

Reference Type BACKGROUND
PMID: 27943336 (View on PubMed)

de Beijer RP, de Graaf C, van Weert A, van Leeuwen TG, Aalders MCG, van Dam A. Identification and detection of protein markers to differentiate between forensically relevant body fluids. Forensic Sci Int. 2018 Sep;290:196-206. doi: 10.1016/j.forsciint.2018.07.013. Epub 2018 Jul 24.

Reference Type BACKGROUND
PMID: 30071450 (View on PubMed)

Other Identifiers

Review additional registry numbers or institutional identifiers associated with this trial.

RECHMPL23_0098

Identifier Type: -

Identifier Source: org_study_id