Mass spectrometry-based methods for the detection of circulating histones h3 and h2b in plasma from sepsis or septic shock (ss) patients
AssigneesCentro de Investigación Biomédica en Red (CIBER-ISCIII), Fundación INCLIVA, Universitat De Valencia Estudi General
InventorsJosé Luis GARCÍA GIMÉNEZ, Carlos ROMÁ MATEO, Federico V. PALLARDÓ CALATAYUD
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Abstract
The present invention proposes a novel method for the detection of circulating histones H3 and H2B in plasma from sepsis or septic shock (SS) patients, based on mass spectrometry-based methods, in particular based on multiple reaction monitoring targeted mass spectrometry (MRM-MS). Such methods allow quantification of histones using an internal standard and show strong specificity and sensitivity values.
Claims12
A mass spectrometry-based method for diagnosing or detecting sepsis or septic shock in a human subject comprising the steps of:
- measuring the level or concentration of at least circulating histone H3 in one or more biological samples consisting of blood, serum or plasma isolated from the subject by using a mass spectrometer;
- and comparing the level or concentration of said circulating histone from the one or more biological samples of the subject suspected of suffering from sepsis or septic shock with a reference value or with the level or concentration of said circulating histone H3 from a biological sample consisting of blood, serum or plasma of a normal subject, wherein the normal subject is a healthy subject not suffering from sepsis or septic shock, wherein the level or concentration of circulating histone H3 is equivalent to the amount of peptide SEQ ID NO 1 (STELLIR) present in the biological sample after trypsin digestion or treatment, and wherein to measure the level or concentration of circulating histone H3 in the biological sample the averaged ratio of SEQ ID NO 1 is obtained, and wherein an increased in the level or concentration of at least circulating histone H3, is indicative of sepsis or septic shock.
A mass spectrometry-based method for predicting the clinical evolution of sepsis or septic shock in a human subject comprising the steps of:
- measuring the level or concentration of at least circulating histone H3 in one or more biological samples consisting of blood, serum or plasma isolated from the subject by using a mass spectrometer;
- and comparing the level or concentration of said circulating histone H3 from the one or more biological samples of the subject suspected of suffering from sepsis or septic shock with a reference value, wherein the level or concentration of circulating histone H3 is equivalent to the amount of peptide SEQ ID NO 1 (STELLIR) present in the biological sample after trypsin digestion or treatment, and wherein to measure the level or concentration of circulating histone H3 in the biological sample the averaged ratio of SEQ ID NO 1 is obtained, wherein an increased in the level or concentration of at least circulating histone H3, is indicative of a negative clinical evolution, and wherein the clinical evolution refers to the overall survival of the patient.
A method for the differential diagnosis of sepsis from septic shock in a human subject suspected of suffering from any of these pathologies, wherein the method comprises the steps of:
- measuring the level or concentration of at least circulating histones H3 and H2B in one or more biological samples consisting of blood, serum or plasma isolated from the subject by using a mass spectrometer;
- and comparing the level or concentration of said circulating histones from the one or more biological samples of the subject suspected of suffering from sepsis or septic shock with a reference value, wherein the level or concentration of circulating histone H3 is equivalent to the amount of peptide SEQ ID NO 1 (STELLIR) present in the biological sample after trypsin digestion or treatment, and to measure the level or concentration of circulating histone H3 in the biological sample the averaged ratio of SEQ ID NO 1 is obtained, and wherein an increased in the level or concentration of at least circulating histone H2B is indicative of septic shock.
A kit which comprises:
- a peptide of SEQ ID NO 1, a peptide of SEQ ID NO 2, or both, wherein these peptides are synthesized with at least one or more heavy amino acids and wherein said peptides are labelled in the C-terminal amino acid residue with a tag, wherein the tag is detectable by mass spectrometry.
12-15. (canceled)
Dependencies are inferred from the claim text.