Method of neonatal serological diagnosis
AssigneesInstitut De Recherche Pour Le Developpement (Ird), Centre National de la Recherche Scientifique (CNRS), Universite Paris Descartes
InventorsFlorence Migot-Nabias, Celia Dechavanne, Francois Guillonneau, Jean-Michel Dugoujon, Marie-Paule Lefranc
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Abstract
The present invention provides a method for determining the amino acid polymorphisms of heavy gamma chain of immunoglobulins G by a proteomic approach. This method distinguishes the immunoglobulins of the mother and those of the newborn in a blood sample obtained during the first months of the child's life. The invention also relates to the use of this method in the early diagnosis of vertically transmitted diseases in the neonate. The invention also provides peptides distinctive of G3m and IGHG3 alleles, and a kit comprising said peptides.
Claims25
1-21. (canceled)
In vitro method for determining the alleles of immunoglobulins G3 (IgG3s) of an individual, the method comprising steps of:
- isolating the IgG3s present in the blood sample from the individual;
- submitting the isolated IgG3s to an enzymatic digestion in order to obtain a mixture of proteotypic peptides of IgG3;
- and
- detecting, by mass spectrometry, among the mixture of proteotypic peptides of IgG3, the presence of at least one peptide distinctive of G3m and/or IGHG3 alleles in order to determine the alleles of the IgG3s present in the blood sample from the individual.
In vitro method for detecting antibodies of a newborn, comprising steps of:
- isolating the IgG3s present in the blood sample obtained from the newborn, the blood sample comprising IgG3s of the newborn and maternal IgG3s transmitted to the newborn during pregnancy;
- submitting the isolated IgG3s to an enzymatic digestion in order to obtain a mixture of proteotypic peptides of IgG3;
- detecting, by mass spectrometry, the presence of at least one peptide distinctive of G3m and/or IGHG3 alleles in order to determine the G3m and IGHG3 alleles of the IgG3s present in the blood sample obtained from the newborn;
- comparing the G3m and IGHG3 alleles of the IgG3s present in the blood sample obtained from the newborn to the G3m and IGHG3 alleles of the IgG3s present in a blood sample obtained from the mother;
- and
- detecting, and optionally quantifying, the IgG3s of the newborn.
In vitro method for diagnosing, in a newborn, an infectious disease caused by a pathogen, the method comprising steps of:
- isolating the pathogen-specific IgG3s present in the blood sample obtained from the newborn, the blood sample comprising pathogen-specific IgG3s of the newborn and pathogen-specific IgG3s of maternal origin transmitted to the newborn during pregnancy;
- submitting the isolated pathogen-specific IgG3s to an enzymatic digestion in order to obtain a mixture of proteotypic peptides of IgG3;
- and
- detecting, by mass spectrometry, among the mixture of proteotypic peptides of IgG3, the presence of at least one peptide distinctive of G3m and/or IGHG3 alleles in order to determine the G3m and IGHG3 alleles of the pathogen-specific IgG3s present in the blood sample obtained from the newborn;
- comparing the G3m and IGHG3 alleles of the pathogen-specific IgG3s present in the blood sample obtained from the newborn to the G3m and IGHG3 alleles of the IgG3s present in a blood sample obtained from the mother;
- and
- detecting, and optionally quantifying, the pathogen-specific IgG3s of the newborn.
Kit for the determination of the alleles of the IgG3s of an individual or for the detection of the IgG3s of a newborn or for the diagnosis of a vertically transmitted disease in a newborn, the kit comprising a plurality of peptides distinctive of G3m and/or IGHG3 alleles belonging to the group consisting of the peptides of SEQ ID NOs:
- 1 to 32 and any combination thereof, wherein the peptides are intended to be used for calibrating the mass spectrometer used in the method.
Dependencies are inferred from the claim text.