EP4100744A1 - Diagnose von kongenitaler zytomegalievirusinfektion - Google Patents

Diagnose von kongenitaler zytomegalievirusinfektion

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Publication number
EP4100744A1
EP4100744A1 EP21750486.9A EP21750486A EP4100744A1 EP 4100744 A1 EP4100744 A1 EP 4100744A1 EP 21750486 A EP21750486 A EP 21750486A EP 4100744 A1 EP4100744 A1 EP 4100744A1
Authority
EP
European Patent Office
Prior art keywords
level
ccmv
symptomatic
biomarker
biological sample
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP21750486.9A
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English (en)
French (fr)
Other versions
EP4100744A4 (de
Inventor
Dana WOLF
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hadasit Medical Research Services and Development Co
Original Assignee
Hadasit Medical Research Services and Development Co
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Hadasit Medical Research Services and Development Co filed Critical Hadasit Medical Research Services and Development Co
Publication of EP4100744A1 publication Critical patent/EP4100744A1/de
Publication of EP4100744A4 publication Critical patent/EP4100744A4/de
Pending legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G01N33/56994—Herpetoviridae, e.g. cytomegalovirus, Epstein-Barr virus
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/60—Complex ways of combining multiple protein biomarkers for diagnosis

Definitions

  • the present invention relates to biomarkers allowing the diagnosis of symptomatic congenital cytomegalovirus (CMV) infection and in particular to differentiate between symptomatic and asymptomatic infected fetuses, to methods of diagnosis of CMV using said biomarkers, diagnostic kits comprising thereof and use of the kits.
  • CMV congenital cytomegalovirus
  • Congenital cytomegalovirus (cCMV) infection resulting from intrauterine transmission, is the most common congenital infection, affecting an average of 0.64% of live births worldwide.
  • Primary human CMV infection during gestation poses -30-40% risk of intrauterine transmission, whereas recurrent infection is associated with a lower transmission rate.
  • cCMV disease develops in -25% of the infected children, and can present as sensorineural hearing loss (SNHL) and a wide range of neurodevelopmental disabilities, brain anomalies, and intrauterine growth restriction.
  • SNHL sensorineural hearing loss
  • amniocentesis with CMV qPCR testing in amniotic fluid is now routinely utilized for the prenatal diagnosis of fetal infection, typically after the 20* week of pregnancy.
  • positive results in amniotic fluid reliably identify fetal infection, they do not distinguish between symptomatic and asymptomatic fetuses/neonate.
  • Prenatal prediction of fetal disease is currently limited, bang mostly based on fetal imaging (by ultrasound or MRI) which is compromised by suboptimal predictive values, especially during early- and mid-gestation, when timely decisions about medical intervention are highly needed.
  • the present invention provides that based on the measurement of concentration(s) of one or more biomarkers as defined in the present application, it is possible to unambiguously diagnose fetus infected with cytomegalovirus and even more, distinguishing between symptomatic and asymptomatic infected fetuses.
  • the present invention provides a method of diagnosing a congenital cytomegalovirus (cCMV) infection, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; and (u) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection, wherein the at least one biomarker is selected from a biomarker of group A, Group B, Group C and Group D, wherein :
  • Group A comprises biomarkers: Cathepsin B (CTSB), Signal-regulatory protein beta- l(SIRPBl), Galectin-3-binding protein (LGALS3BPX Agrin (AGRN), Vascular cell adhesion protein 1 (VCAM1X Coactosin-like protein (COTL1X CDS antigen-like (CD5LX ATP synthase subunit beta, mitochondrial (ATP5BX Keratin, type II cytoskeletal 6A (KRT6AX Secreted and transmembrane protein 1 (SECTM1X Fatty acid-binding protein, heart (FABP3X Cal cy cl in (S100A6X (SH3 domain-binding glutamic acid-rich-like protein (SH3BGRLX Interferon-stimulated gene 15 (ISG15X Cartilage acidic protein 1 (CRTAC1X Retinoic acid receptor responder protein 2 (RARRES2X Bone marrow stromal antigen 2 (BST2), Beta-2
  • Group D comprises biomarkers CPM, Platelet-derived growth factor receptor beta (PDGFRB), Angiopoietin-related protein 6 (ANGPTL6), Dermokine (DMKN), Secretogranin-1 (CHGB), ADAM9, Growth anest-specific protein 1 (GAS1), Platelet basic protein (PPBP), Platelet glycoprotein lb alpha chain (GP1BA).
  • the biomarkers are protein biomarkers.
  • the biological sample is obtained from a subject selected from a fetus, pregnant woman and neonate.
  • the biological sample is selected from an amniotic fluid, fetal blood, fetal blood from the umbilical cord, placental biopsy, neonate blood and maternal blood or serum.
  • an increase in the level of at least one biomarker of Group A or C, and/or a decrease in the level of at least one biomarker of Group B or D in the biological sample in comparison to their levels in the control or in comparison to a predefined cutoff level is indicative of cCMV infection.
  • the present invention provides a method for diagnosing a symptomatic cCMV.
  • the method of diagnosing according to the present invention allows differentiating between symptomatic and asymptomatic cCMV infected subjects, such as symptomatic and asymptomatic fetuses and/or neonates.
  • an increase in the level of at least one biomarker of Group C and/or a decrease in the level of at least one biomarker of Group D in the biological sample in comparison to their levels in the control or in comparison to a predefined threshold level is indicative of a symptomatic cCMV infection.
  • the present invention provides a method of diagnosing a symptomatic congenital cytomegalovirus (symptomatic cCMV) in a fetus, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of presence of the symptomatic cCMV infection, wherein the at least one biomarker is selected from the biomarkers of group Group C and Group D, wherein: Group C comprises biomarkers RARRES2, LGALS3BP, GPNMB, Nesprin-1 (SYNE1), Myosin- 14 (MYH14), Carboxypeptidase A2 (CPA2), and Secretoglobin family 3A member 2 (SCGB3A2); and Group D comprises biomarkers CPM, Platelet -derived growth
  • the biomarkers are protein biomarkers.
  • the biological sample is obtained from a subject selected from a fetus and pregnant woman.
  • the biological sample is selected from an amniotic fluid, fetal blood, plasma or serum, fetal blood, plasma or serum from the umbilical cord, placental biopsy, and maternal blood or serum.
  • the biological sample is an amniotic fluid.
  • an increase in the level of RARRES2 in the biological sample in comparison to its level in the control or in comparison to a predefined threshold level is indicative of symptomatic cCMV infection.
  • an increase in the level of LG AL S3 BP in the biological sample in comparison to its level in the control or in comparison to a predefined threshold level is indicative of a symptomatic cCMV infection.
  • an increase in the level of GPNMB in the biological sample in comparison to its level in the control or in comparison to a predefined threshold level is indicative of a symptomatic cCMV infection.
  • the method of diagnosis comprises determining levels of at least two, at least 3, at least 4 or at least 5 biomarkers in the biological sample.
  • the method of diagnosis comprises determining levels of 2, 3, 4, 5 or 6 biomarkers in the biological sample.
  • an increase in the levels of (i) RARRES2, (ii) LGALS3BP or (iii) both RARRES2 and LGALS3BP in the biological sample in comparison to their levels in the control or in comparison to their predefined threshold levels is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARRES2 and of LGALS3BP, and (i) an increase in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, and SCGB3A2 or (ii) a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in comparison to their levels in the control or in comparison to their predefined threshold levels is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARRES2, LGALS3BP, and GPNMB in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • the biological sample is an amniotic fluid.
  • the biological sample is a maternal serum.
  • the diagnosis is performed before week 36 of the pregnancy.
  • the method further comprises a step of diagnosing whether the subject, e. g a fetus, is infected with CMV.
  • the method further comprises differentiating between symptomatic and asymptomatic CMV infection.
  • the present invention provides a method of diagnosing an asymptomatic congenital cytomegalovirus in a fetus, wherein the method comprises (i) diagnosing whether the fetus is infected with CMV and (ii) diagnosing whether the fetus has a symptomatic CMV by the method of the present invention, wherein lack of symptomatic CMV in the infected fetus is indicative of presence of asymptomatic cCMV.
  • the present invention provides a method for recommending a medical intervention, the method comprises diagnosing a congenital cytomegalovirus using the methods of diagnosis according to the present invention, and recommending the medical intervention based on the obtained results.
  • the congenital cytomegalovirus is a symptomatic cCMV.
  • the congenital cytomegalovirus is an asymptomatic cCMV.
  • the present invention provides a method for recommending a medical intervention comprising detecting a symptomatic or asymptomatic congenital cytomegalovirus in fetus according to the methods of the present invention and recommending the medical intervention based on the obtained results.
  • the present invention provides a method of executing a medical intervention in case of existence of a congenital CMV such symptomatic cCMV comprising diagnosing the cCMV according to the methods of the present invention and effecting the medical intervention.
  • the medical intervention is a treatment with an anti-CMV active agent.
  • the medical intervention is a termination of pregnancy.
  • the present invention provides a method of treating a congenital cytomegalovirus in a fetus comprising diagnosing the symptomatic or asymptomatic cCMV in the fetus according to the methods of the present invention and administering to the fetus or the mother an anti-CMV active agent.
  • the congenital cytomegalovirus is a symptomatic CMV.
  • the congenital cytomegalovirus is an asymptomatic CMV.
  • the present invention provides a kit comprising means for determining a level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group A, Group B, Group C and Group D, and instructions for use.
  • the kit comprises instructions for diagnosing a congenital cytomegalovirus (cCMV) infection.
  • the kit comprises instructions for diagnosing a symptomatic cCMV.
  • the kit comprises instructions for diagnosing an asymptomatic cCMV.
  • the kit provides means for determining the levels of a biomarker selected from RARRES2, LGALS3BP, GPNMB, and a combination thereof.
  • the present invention provides a kit comprising means for determining levels of at least two biomarker in a biological sample, wherein the at least two biomarkers are selected from biomarkers of Group C and Group D, and instructions for use of said kit.
  • the kit comprises means for determining the levels of RARRES2 and LGALS3BP, RARRES2 and GPNMB or LGALS3BP and GPNMB or of RARRES2, LGALS3BP and GPNMB.
  • the kit further comprises means for determining the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA.
  • the means of the kit allow detecting levels of a plurality of biomarkers.
  • the kit comprises all required means and information for detecting the levels of the biomarkers in the biological samples and for comparing the detected levels to their levels in the control or to their threshold levels for diagnosing the cCMV and/or symptomatic cCMV.
  • the kit of the present invention is for diagnosing cCMV.
  • the kit of the present invention is for diagnosing symptomatic cCMV.
  • the present invention provides use of a kit in diagnosis a congenital CMV, wherein the kit comprises means for determining a level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group A, Group B, Group C and Group D.
  • the use comprises diagnosing a symptomatic cCMV
  • the use comprises diagnosing an asymptomatic cCMV.
  • the use comprises determining the levels of RARRES2 and/or LGALS3BP in a biological sample, and optionally further determining the levels of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, GP1BA, and comparing the determined levels to the threshold levels, thereby diagnosing the cCMV and/or symptomatic or asymptomatic cCMV.
  • biomarker selected from SYNE1, MYH14, GPNMB, CPA2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, GP1BA
  • Fig. 1 shows a heatmap showing the proteins differing between CMV-positive (yes) and C MV -negative (no) amniotic fluid samples.
  • the heatmap was drawn using normalized values, after scaling per protein over all drawn samples.
  • Fig. 2A shows a heatmap showing amniotic fluid proteins differing between severely symptomatic (sick) and asymptomatic (not sick).
  • Fig.2B shows a heatmap showing the same set of proteins shown in Fig. 2A in severely symptomatic (sick) asymptomatic (not sick) and CMV-negative amniotic fluid samples. Heatmaps were drawn using normalized values, after scaling per protein over all drawn samples.
  • Fig. 3A shows the distribution and median values of RARRES2 protein concentrations, as measured by ELISA immunoassay, in amniotic fluid samples from symptomatic (sickX asymptomatic (not sickX and CMV-negative (uninfected) cCMV cases.
  • FIG. 3B shows RARRES2 protein concentrations, as measured by ELISA, in individual amniotic fluid samples from symptomatic (sick; blackX asymptomatic (not sick; grey), and CMV-negative (uninfected; white) cases. Samples which had been pre-analyzed by proteome analysis (the discovery panel) are indicated by + sign.
  • FIG. 4A shows the distribution and median values of LGALS3BP protein concentrations, as measured by ELISA, in amniotic fluid samples from symptomatic (sick), asymptomatic (not sick), and CMV-negative (uninfected) cCMV cases.
  • Fig. 4B shows LGALS3BP protein concentrations, as measured by ELISA, shown in individual amniotic fluid samples from symptomatic (sick; black), asymptomatic (not sick; grey), and CMV- negative (uninfected, white) cases. Samples which had been pie-analyzed by proteome analysis (the discovery panel) are indicated by “+” sign.
  • the present invention is based on an unexpected observation that a set of biomarkers, i .e. proteins, secreted into the amniotic fluid, may be useful to unambiguously diagnose fetal infected with cytomegalovirus. Even more surprising was the finding that several specific biomarkers allow determining whether the fetus has/will develop a symptomatic congenital CMV. In other word, the methods of the present invention allow differentiating between symptomatic and asymptomatic infected fetuses. Currently this differentiation is often made by imaging at a later stage and usually does not provide a robust differentiation.
  • Determination whether the fetus is infected by CMV is usually made by measuring viral load in am ni otic fluid and usually is unambiguous. However, diagnosing whether the fetus is expected to develop symptoms of CMV will allow to provide much more specific recommendations and will reduce the number of non-necessary terminations of pregnancies. As shown in the results, the present invention allows differentiating fetus having symptomatic and asymptomatic CMV with sensitivity, specificity, and positive and predictive values of above 95.0% each (referring to RARRES2 and LGALS3BP proteins).
  • detecting RARRES2 in amniotic fluids in concentration above 40 ng/ml allows diagnosing a symptomatic cCMV with sensitivity of 95.2%, specificity of 92.9%, positive predictive value of 90.9%, and negative predictive value of 96.3% %.
  • detection of LGALS3BP in amniotic fluid above 2475 ng/ml allowed a reliable prenatal disease prediction with sensitivity of 90.5%, specificity of 96.4%, positive predictive value of 95.0%, and negative predictive value of 93.1%.
  • the present invention provides a method of diagnosing a congenital cytomegalovirus (cCMV) infection, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection, wherein the at least one biomarker is selected from Cathepsin B (CTSB), Signal- regulatory protein beta-l(SIRPBl), Galectin-3-binding protein (LGALS3BP), Agrin (AGRN) » Vascular cell adhesion protein 1 (VCAM1), Coactosin-like protein (COTL1), CDS antigen-like (CD5L), ATP synthase subunit beta, mitochondrial (ATP5B), Keratin, type ⁇ cytoskeletal 6A (KRT6A), Secreted and
  • CTSB Cat
  • Group A of biomarkers comprises biomarkers CTSB, SIRPB1, LGALS3BP, AGRN, VCAM1, COTL1, CD5L, ATP5B, KRT6A, SECTM1, FABP3, S100A6, SH3BGRL, ISG15, CRTAC1, RARRES2, BST2, B2M, EPYC, DKK1, HLA-C, CPQ, F5, GPNMB, LAMAS, CA3, and DES
  • Group B of biomarkers comprises biomarkers MEGF8, GALNT7, FAT4, LPHN2, AFM, MASP1, SOD3, GALNT2, LSR, HGF, PRG2, QSOX1, ERVMER34-1, PAPPA2, GOT1, RDX IGFBP5, AHNAK, ADAM9, LAMP2, CPM, SBSN, CAST, SPINKS, TNMD, CDH15, KLKB1, PRSS
  • Group B of biomarkers comprises biomarkers Disintegrin and metalloproteinase domain-containing protein 9 (ADAM9), Multiple epidermal growth factor-like domains protein 8 (MEGF8), N-acetylgalactosaminyl transferase 7 (GALNT7), Protocadherin Fat 4 (FAT4), (Latrophilin-2 (LPHN2), (Afamin (AFM), Mannan-binding lectin serine protease 1 (MASP1), Extracellular superoxide dismutase [Cu-Zn] (SOD3), Polypeptide N- acetylgalactosaminyltransferase 2 (GALNT2), Lipdysis-stimulated lipoprotein receptor (LSR), Hepatocyte growth factor (HGF), Bone marrow proteoglycan (PRG2), Sulfhydryl oxidase 1 (QSOX1), Endogenous retrovirus group MER34 member 1 Env poly
  • GAT1 Radixin (RDX), Insulin-like growth factor-binding protein 5 (IGFBP5), Desmoyokin (AHNAK), Ly sosom e-associ ated membrane glycoprotein 2 (LAMP2), Carboxypeptidase M (CPM), Suprabasin (SBSN), Calpastatin (CAST), Serine protease inhibitor Kazal-type 5 (SPINKS), Tenomodulin (TNMD), Cadherin-15 (CDH15), Plasma kallikrein (KLKB1), Prostasin (PRSS8), Collagen alpha-l(m) chain (COL3A1X and Collagen alpha-2(I) chain (COL1 A2).
  • Group C of biomarkers comprises biomarkers SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, and SCGB3A2
  • Group D of biomarkers comprises biomarkers CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA.
  • the biomarkers are protein biomarkers.
  • the present invention provides a method of diagnosing a congenital cytomegalovirus (cCM V) infection, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the sample to its level in a control, wherein the difference between the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection, wherein the at least one biomarker is selected from the biomarkers of group A, Group B, Group C and Group D, as defined hereinabove.
  • cCM V congenital cytomegalovirus
  • the biomarkers i.e. the proteins are human proteins.
  • the proteins are fetal proteins.
  • the biological sample is obtained from a subject.
  • subject refers to a human subject.
  • the subject is selected from a fetus, pregnant woman and neonate.
  • biological sample refers to a sample obtained from a subject, including a sample of a biological tissue or fluid origin obtained in vivo or in vitro.
  • Biological samples can be, without limitation, body fluid (e g., blood, blood plasma, serum), organs, tissues, fractions and cells isolated from the subject/patient.
  • Biological samples also may include sections of the biological sample including tissues (e g., sectional portions of an organ or tissue).
  • Biological samples may be dispersed in solution or may be immobilized on a solid support, such as in blots, assays, arrays, glass slides, microtiter, or ELISA plates.
  • Biological samples may be processed in any known method before their use in the methods of the present invention.
  • the biological sample is an amniotic fluid.
  • the biological sample is selected from an amniotic fluid, fetal blood, fetal blood serum, fetal plasma, blood, serum or plasma from the umbilical cord, placental biopsy, neonate blood, neonate dried blood spot and maternal blood or serum.
  • the biological sample is selected from the group consisting of breast milk, maternal blood, maternal urine, maternal saliva, fetal blood, fetal blood from the umbilical cord, postnatal infant urine, blood, saliva, a uterine biopsy sample, and a placental biopsy.
  • the biological sample is fetal blood.
  • the biological sample is maternal blood.
  • the biological sample is neonate blood.
  • the term blood encompasses the term “serum” and may be replaced by it.
  • the biological sample is a maternal serum.
  • the biological sample is a fetal or neonatal serum.
  • the biological sample is neonatal dried blood spot.
  • the fetal marker is measured in the biological sample obtained from the mother such as maternal blood, serum or milk.
  • marker and “biomarker” are used interchangeably and refer to a molecule (typically a protein, nucleic acid, caifrohydrate, or lipid) that is differentially expressed in the cell, differentially expressed on the surface of an infected cell, differentially phosphorylated, or differentially secreted by an infected cell in comparison to a normal cell or in a paracrine fashion by neighboring uninfected cells, and which is useful for the diagnosis of congenital CMV infection or symptomatic congenital CMV infection for providing a prognosis for birth defects, and for preferential targeting of a pharmacological agent to an infected fetus or individual.
  • a molecule typically a protein, nucleic acid, caifrohydrate, or lipid
  • the biomarker is a protein
  • such markers are proteins that are secreted by an infected cell in comparison to a normal cell, for instance, 1-fold secretion, 2-fold secretion, 3-fold secretion or more in comparison to a normal cell.
  • biomarkers are molecules that are underexpressed in an infected cell in comparison to a normal cell.
  • the marker is a nucleic acid molecule.
  • the marker is a nucleic acid molecule such as DNA or RNA molecule correlating to the amount of a protein.
  • a marker can be a molecule that is inappropriately synthesized in the infected cell, for instance, a molecule that contains deletions, additions or mutations in comparison to the molecule expressed on a normal cell.
  • a marker can also be a molecule that is inappropriately processed in infected cells, for instance, a molecule that is secreted, protedytically processed or subject to post-translational modification (e g., phosphorylation, glycosylation) in comparison to the molecule expressed on a normal cell.
  • the term "biomarker” as used herein refers to a protein that is differentially present in a biological sample, such as a protein that is differentially excreted into amniotic fluid. According to some embodiments, the biomarkers, such as proteins, are present in biological samples, whereas the presence of the marker above or below a particular amount, referred also as a threshold or cutoff value, is indicative of a diagnosed condition.
  • cytomegalovirus and “CMV” are uses herein interchangeably and refer to a genus of viruses of herpes virus as known in the art, more specifically to CMV infecting humans.
  • congenital CMV infection and “cCMV” are used herein interchangeably and refer to in utero transmitted CMV infection. The term refers also to an infection in fetuses, neonates and children that were infected in utero.
  • the congenital CMV may be symptomatic and asymptomatic.
  • symptomatic cCMV and “symptomatic congenital CMV”, as used in the present invention and claims, refer to the condition in which the infected subject demonstrates a moderate to severe cCMV disease in utero and/or after birth. This term contemplates the expectancy that the fetus will develop a moderate to severe cCMV disease in utero and/or after birth.
  • asymptomatic cCMV asymptomatic congenital CMV
  • non-symptomatic cCMV refer to the condition in which the infected subject does not demonstrate or is not expected to demonstrate signs of cCMV disease in utero and does not develop or is not expected to develop symptoms/signs of a disease caused by CMV for at least 1 year after birth.
  • infected refers to subject that is infected with CMV virus and may be symptomatic and asymptomatic.
  • symptomatic cCMV results is sensorineural hearing loss (SNHL), a wide range of neurodevelopmental disabilities, brain anomalies, and intrauterine growth restriction.
  • moderate to severely symptomatic cCMV disease includes: Multiple manifestations attributable to cCMV infection, such as: thrombocytopenia, petechiae, hepatomegaly, splenomegaly, intrauterine growth restriction, hepatitis (raised transaminases or bilirubin), and/or
  • central nervous system involvement such as: microcephaly, radiographic abnormalities consistent with cytomegalovirus central nervous system disease (ventriculomegaly, intracerebral calcifications, periventricular echogenicity, cortical or cerebellar malformations), abnormal cerebrospinal fluid indices for age, chorioretinitis, sensorineural hearing loss, and the detection of cytomegalovirus DNA in cerebrospinal fluid (Rawlinson et al., Lancet Infect Dis 2017; 17:el 77-el 88.) [0031] As used herein, the term “lever refers to the amount or concentration of a biomarker
  • biomarker e g. protein
  • ELISA ELISA-binding protein-binding protein
  • Methods for determining the levels of the biomarker correspond to the type of biomarkers.
  • nucleic acid biomarkers such as DNA or RNA may be determined by any known appropriate method.
  • control refers to biological samples from a particular predefined group of subjects.
  • control refers to the biological samples obtained from n on-infected subjects, from infected n on-symptomatic or to combination thereof.
  • level in the control’’ with respect to a biomarker has the meaning of the level, e g amount or concentration, of the biomarker in biological samples of the predefined group of subjects, as defined above.
  • the control comprises samples of non -infected fetuses, infected n on-symptomatic fetuses, or both.
  • diagnosis also refers, in some embodiments, to screening Screening for infection, in some embodiments, can lead to earlier diagnosis in specific cases.
  • the term refers also to detecting congenital CMV infection.
  • the term in some embodiments refers to distinguishing between symptomatic and asymptomatic congenital CMV infection.
  • the term encompasses assessing whether the subject has a symptomatic cCMV infection.
  • the term also encompasses assessing whether the subject will develop a symptomatic cCMV infection.
  • the term “providing a prognosis” refers to providing a prediction of the probable course and outcome of the diagnosis.
  • diagnosis cCMV have the meaning of predicting whether the subject has or will be infected by cCMV.
  • diagnosing symptomatic cCMV have the meaning of predicting whether the subject has or will be infected by symptomatic cCMV. As stated above, the prediction is not always 100%. Thus, the term “diagnosing symptomatic cCMV” may have the meaning predicting that the subject has or will have symptomatic cCMV with a probability of 95%, 90%, 85%, 80%, 75%, 70%, or from 70 to 95%. According to some embodiments, the sensitivity of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%. According to some embodiments, the specificity in diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the positive predictive value of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the negative predictive value of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • sensitivity refers to the ratio of positive samples correctly recognized as such by carrying out the assay of interest and the total number of samples examined. In other words, an assay has 100% sensitivity if all samples from infected patients give a positive result, i.e. there are no false negative results.
  • positive predictive value refers to the probability of the disease being present, among those with positive diagnostic test results.
  • negative predictive value refers to the probability that the disease was absent, among those whose diagnostic test results were negative.
  • the diagnosed subject is a fetus. Therefore, the present invention provides a method of prenatal diagnosis of a congenital CMV in fetus. According to some embodiments, the present invention provides a method of prenatal diagnosis of a symptomatic congenital CMV in fetus.
  • an increase in the level of at least one biomarker of Group A in the biological sample in comparison to its levels in the control is indicative of cCMV infection.
  • an increase in the level of at least one biomarker of Group A in the biological sample above a particular ratio is indicative of cCMV infection.
  • a ratio of above 1.1.1 between the level of at least one biomarker of Group A in the biological sample to its levels in the control is indicative of cCMV infection.
  • a decrease in the level of at least one biomarker of Group B in the biological sample in comparison to its levels in the control is indicative of cCMV infection.
  • a decrease in the level of at least one biomarker of Group B in the biological sample below a particular threshold is indicative of cCMV infection.
  • a ratio of above 1.1:1 between the level of the at least one biomarker in the control to the level of at least one biomarker of Group B in the biological sample is indicative of cCMV infection.
  • the method of diagnosis comprises determining the level of more than one biomarker.
  • an increase in the levels of 2 or more biomarkers of Group A in the biological sample in comparison to their levels in the control is indicative of cCMV infection.
  • an increase in the level of 3 or more biomarkers of Group A in the biological sample in comparison to their levels in the control is indicative of cCMV infection.
  • a decrease in the level of 2 or more biomarkers of Group B in the biological sample in comparison to their levels in the control is indicative of cCMV infection.
  • a decrease in the level of 3 or more biomarkers of Group B in the biological sample in comparison to their levels in the control is indicative of cCMV infection.
  • the biomarker is selected from RARRES2, LG AL S3 BP, GPNMB and a combination thereof.
  • an increase in the level of a biomarker selected from RARRES2, LGALS3BP, GPNMB and a combination thereof in comparison to their levels in the control is indicative of presence of cCMV infection in the subject.
  • the biomarker is selected from SYNE1, ANGPTL6, GAS1, CHGB, ADAM9, and any combination thereof.
  • an increase in the level of SYNE1 and/or a decrease in the level of ANGPTL6, GAS 1 , CHGB, or ADAM9 in the biological sample in comparison to their levels in the control is indicative of presence of cCMV infection in the subject.
  • a difference in the levels of the biomarker in maternal blood and in the control is indicative of a congenital CMV infection in fetus.
  • a difference in the levels of the biomarker in amniotic fluid and in the control is indicative of a congenital CMV infection in fetus.
  • the control comprises samples from n on-infected subjects.
  • a level of the biomarker in the control refers to a level of the biomarker in biological samples of n on-infected subjects.
  • the subject is fetus.
  • the subject is a pregnant woman, h is clear that the biological sample and the control refer correspond one to another.
  • a level of the biomarker in the control refers to a level of the biomarker in amniotic fluid of n on-infected fetuses.
  • diagnosing comprises diagnosing of a symptomatic cCMV.
  • diagnosing of the symptomatic cCMV comprises differentiating between symptomatic and asymptomatic cCMV.
  • the present invention provides a method of diagnosing a symptomatic congenital cytomegalovirus (symptomatic cCMV) in a subject, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of presence of a symptomatic cCMV infection in the subject, wherein the at least one biomarker is selected from the biomarkers of group Group C and Group D, wherein: Group C comprises biomarkers RARRES2, LGALS3BP, GPNMB, Nesprin-1 (SYNE1), Myosin-14 (MYH14), Caiboxypeptidase A2 (CPA2), Secretoglobin family 3A member 2 (SCGB3A2), and Group D comprises biomarkers CPM, Platelet-
  • the biomarkers are protein biomarkers.
  • the biological sample is obtained from a subject selected from a fetus, pregnant woman and neonate.
  • the biological sample is an amniotic fluid.
  • the biological sample is selected from amniotic fluid, fetal blood or serum, fetal blood or serum from the umbilical cord, placental biopsy, neonate blood or serum and maternal blood or serum.
  • the biological sample is an amniotic fluid.
  • the sample is a sample of an infected subject.
  • the sample is a sample of an infected fetus.
  • diagnosing the presence of CMV infection may be according to any known method, e g.
  • RARRES2 refers to human Retinoic Acid Receptor Responder 2 protein having accession number Q99969.
  • LGALS3BP refers to a human Galectin-3-binding protein having accession number Q08380.
  • GPNMB refers to a human transmembrane glycoprotein NMB having accession number Q 14956.
  • an increase in the level of 1, 2, 3, 4, 5, 6, or 7 of the biomarkers of group C in comparison to their levels in the control is indicative of cCMV infection in the subject.
  • a decrease in the level of 1, 2, 3, 4, 5, 6, 7, 8 or 9 of the biomarkers of group D in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • an increase in the level of 1, 2, 3, 4, 5, 6, or 7 of the biomarkers of group C and a decrease in the level of 1, 2, 3, 4, 5, 6, 7, 8, or 9 of the biomarkers of group D in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample in comparison to its levels in the control is indicative of a symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker of Group D in the biological sample below a particular threshold is indicative of a symptomatic cCMV infection.
  • a ratio of above 1.1:1 between the level of the at least one biomarker in the control to the level of the least one biomarker of D in the biological sample is indicative of a symptomatic cCMV infection.
  • an increase in the level of at least one biomarker of Group C and a decrease in the level of at least one biomarker of Group D in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • the biological sample is an amniotic fluid.
  • the present invention provides a method of diagnosing a symptomatic congenital cytomegalovirus (cCMV) infection, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a symptomatic congenital CMV infection, wherein the at least one biomarker is selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA.
  • cCMV congenital cytomegalovirus
  • the control comprises samples from the infected n on-symptomatic, n on-infected subject or both.
  • a level of the biomarker in the control refers to a level of the biomarker in biological samples of infected non- symptomatic, in the samples of non -infected subjects, or in combination thereof.
  • the subject is a fetus.
  • the subject is a pregnant woman.
  • the level is an amount or a concentration of the biomarker in the sample.
  • the biological sample is an amniotic fluid.
  • an increase in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, SCGB3A2 in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of more than 1.5, 2, 2.5 or 3 folds in the level of the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of more than 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% 60%, 70%, 80 or 100% in the level of the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, SCGB3 A2 in the biological sample above a particular value or threshold as defined based on the control is indicative of a symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA more than 10%, more than 15%, more than 20% or more than 25 % in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample below a particular value or threshold as defined based on the control is indicative of a symptomatic cCMV infection.
  • comparison to the level in a control may be performed by defining a particular threshold above which or below which the level of the marker is considered as indicative to cCMV, and in particular indicative to symptomatic cCMV.
  • the term “comparing the level of the at least one biomarker from the biological sample to its level in a control” and alike may be replaced by “comparing the level of the at least one biomarker to a threshold”.
  • the term “an increase in the level of a biomarker X in the biological sample in comparison to their levels in the control” and alike may be replaced by the term “an increase in the level of a biomarker X in the biological sample above a threshold”.
  • the method comprises determining the level of
  • an increase in the level of RARRES2 in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of more than 1.5, 2, 2.5 or 3 folds in the level of RARRES2 in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of from 2 to 20 folds in the level of RARRES2 in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of from 4 to 16, 5 to 15, 8 to 12 folds is indicative of a symptomatic cCMV infection.
  • an increase of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100%, of above 150%, above 200%, above 250%, above 300% in the level of RARRES2 in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of RARRES2 in the biological sample above a particular value as define based on the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of RARRES2 of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100% or more, or from 2 to 20 times above a threshold is indicative of a symptomatic cCMV infection.
  • the threshold is 40 ng/ml of RARRES2 in amniotic fluid, as determined by the methods of the present invention.
  • the threshold is 51 ng/ml.
  • an increase in the level of RARRES2 in the biological sample above 30 ng/ml, above 35 ng/ml or above 40 ng/ml or above 45 ng/ml or above 50 ng/ml or above 51 ng/ml or above 55 ng/ml is indicative of a symptomatic cCMV infection.
  • the threshold for RARRES2 in amniotic fluid is from 20 to 80 ng/ml.
  • the threshold for RARRES2 in amniotic fluid is from 25 to 75, from 30 to 70, or from 35 to 65 ng/ml.
  • the sensitivity of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the specificity in diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the positive predictive value of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the negative predictive value of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the method comprises determining the level of LGALS3BP.
  • an increase in the level of LGALS3BP in the biological sample in comparison its level to the control is indicative of a symptomatic cCMV infection.
  • the increase in the level of LGALS3BP in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of LG AL S3 BP in the biological sample above a particular value as defined based on the control is indicative of a symptomatic cCMV infection.
  • increase of more than 1.5, 2, 2.5 or 3 folds in the level of LGALS3BP in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of from 2 to 20 folds in the level of LGALS3BP in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of from 4 to 16, 5 to 15, 8 to 12 folds is indicative of a symptomatic cCMV infection.
  • an increase of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100%, of above 150%, above 200%, above 250%, above 300% of LGALS3BP in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of LG AL S3 BP of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100% or more, or from 2 to 20 times above a threshold is indicative of a symptomatic cCMV infection.
  • the threshold for is 2475 ngZml of LGALS3BP in amniotic fluid, as determined by the methods of the present invention.
  • an increase in the level of LGALS3BP in the biological sample above 1500 ngZml, above 2000 ng/ml or above 2200 ng/ml or above 2400 ngZml or above 2475 ng/ml or above 2500 ng/ml or above 2700 ng/ml is indicative of a symptomatic cCMV infection.
  • the threshold for LG AL S3 BP in amniotic fluid is from 1000 to 4000 ng/ml.
  • the threshold for LGALS3BP in amniotic fluid is from 1200 to 3800 ng/ml, from 1500 to 3500 ng/ml, from 1700 to 3200 ng/ml, from 2000 to 3000 ng/ml, or from 2200 to 2800 ng/ml.
  • the sensitivity of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the specificity in diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the positive predictive value of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the negative predictive value of diagnosing symptomatic cCMV is from 85 to 99%, from 90 to 98% or from 92 to 96%.
  • the method comprises determining the level of GPNMB.
  • an increase in the level of GPNMB in the biological sample in comparison to its level in the control is indicative of symptomatic cCMV.
  • an increase of more than 1.5, 2, 2.5 or 3 folds in the level of GPNMB in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase of from 2 to 20 folds in the level of GPNMB in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • increase of from 4 to 16, 5 to 15, 8 to 12 folds is indicative of a symptomatic cCMV infection.
  • an increase of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, above 100%, above 150%, above 200%, above 250%, above 300% of GPNMB in the biological sample in comparison to its level in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of GPNMB of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100% or more, or from 2 to 20 times above a threshold is indicative of a symptomatic cCMV infection.
  • the method comprises determining the level of SYNE1.
  • an increase in the level of SYNE1 in the biological sample in comparison to its level in the control is indicative of symptomatic cCMV.
  • an increase in the level of SYNE 1 of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100% or more, or from 2 to 20 times above a threshold is indicative of a symptomatic cCMV infection.
  • the method comprises determining the level of ANGPTL6.
  • a decrease in the level of ANGPTL6 in the biological sample in comparison to its level in the control is indicative of symptomatic cCMV.
  • a decrease in the level of ANGPTL6 of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more in comparison to a threshold is indicative of a symptomatic cCMV infection.
  • the method comprises determining the level of
  • a decrease in the level of GAS1 in the biological sample in comparison to its level in the control is indicative of symptomatic cCMV.
  • a decrease in the level of GAS1 of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more in comparison to a threshold is indicative of a symptomatic cCMV infection.
  • the method comprises determining the level of CHGB.
  • a decrease in the level of CHGB in the biological sample in comparison to its level in the control is indicative of symptomatic cCMV.
  • a decrease in the level of CHGB in the biological sample of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more in comparison to a threshold is indicative of symptomatic cCMV.
  • the method comprises determining the level of ADAM9.
  • a decrease in the level of ADAM9 in the biological sample in comparison to its level in the control is indicative of symptomatic cCMV.
  • a decrease in the level of ADAM9in the biological sample of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more in comparison to a threshold is indicative of symptomatic cCMV.
  • the method comprises determining the level of at least two biomarkers selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample.
  • the method comprises determining the levels of RARRES2 and LGALS3BP.
  • an increase in the levels of RARRES2 and of LGALS3BP in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • an increase of 10% or more, 15% or more, 20% or more, 25 % or more, 50% or more, 75% or more, 100% or more, or from 2 to 20 times above thresholds in the levels of RARRES2 and of LGALS3BP is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARRES2 above 35 ng/ml, 40 ng/ml or above 50 ng/ml and an increase in the levels of LGALS3BP above 2400 ng/ml, above 2450 ng/ml or above 2475 ng/ml is indicative of a symptomatic cCMV infection.
  • the method comprises determining the levels of RARRES2 and LGALS3BP and an increase in the levels of RARRES2 or LGALS3BP in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • the method comprises determining the levels of RARRES2, LGALS3BP and/or GPNMB.
  • an increase in the levels of RARAES2 and of GPNMB in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the levels of LGALS3BP and GPNMB in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARRES2, LGALS3BP or GPNMB is indicative of symptomatic cCMV infection.
  • an increase in the levels of RARRES2, LGALS3BP and GPNMB is indicative of symptomatic cCMV infection.
  • an increase in the levels of RARRES2 and of LGALS3BP, and in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, and SCGB3A2 in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARRES2 and of LG AL S3 BP, and a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARAES2, LGALS3BP and GPNMB in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the levels of RARRES2, LGALS3BP and SYNE1 in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • an increase in the levels of RARRES2, LGALS3BP, GPNMB and SYNE1 in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • CHGB, ADAM9 or a combination thereof in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • the biological sample is amniotic fluid.
  • an increase in the levels of RARRES2 and LGALS3BP and a decrease in the level of ADAM9 in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • an increase in the levels of RARRES2, GPNMB and LGALS3BP and a decrease in the level of a biomarker selected from ANGPTL6, GAS1, CHGB, ADAM9 or a combination thereof in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • the biological sample is amniotic fluid.
  • an increase in the levels of RARRES2, GPNMB and LGALS3BP and a decrease in the level of ADAM9 in the biological sample in comparison to their levels in the control is indicative of symptomatic cCMV infection.
  • an increase in the levels of at least one or at least two or at least three biomarkers selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, and SCGB3A2 in amniotic fluid in comparison to their levels in the control is indicative of a symptomatic cCMV infection in fetus.
  • an increase in the levels of RARRES2 and/or LGALS3BP, and optionally in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, and SCGB3A2 in amniotic fluid in comparison to their levels in the control is indicative of a symptomatic cCMV infection in fetus.
  • the present invention provides method of diagnosing a symptomatic congenital cytomegalovirus (symptomatic cCMV) in a fetus, wherein the method comprises (i) determining a levels of biomarkers RARRES2, LGALS3BP or both in amniotic fluid; and (ii) comparing the levels determined in (i) to their levels in a control, wherein an increase in the level of the biomarker in the biological sample in comparison to their levels in the control is indicative of presence of a symptomatic congenital cCMV infection in fetus.
  • the method comprises (i) determining a levels of biomarkers RARRES2, LGALS3BP or both in amniotic fluid; and (ii) comparing the levels determined in (i) to their levels in a control, wherein an increase in the level of the biomarker in the biological sample in comparison to their levels in the control is indicative of presence of a symptomatic congenital cCMV infection in fetus.
  • the present invention provides a method of diagnosing a symptomatic congenital cytomegalovirus (symptomatic cCMV) in a fetus, wherein the method comprises (i) determining a levels of biomarkers RARRES2, LGALS3BP or both in amniotic fluid, and (ii) comparing the levels determined in (i) predefined thresholds, wherein an increase in the level of the biomarker in the biological sample in comparison to the thresholds is indicative of presence of a symptomatic congenital cCMV infection in fetus.
  • symptomatic congenital cytomegalovirus symptomatic congenital cytomegalovirus
  • the present invention provides a method of diagnosing a symptomatic congenital cytomegalovirus (symptomatic cCMV) in a fetus, wherein the method comprises (i) determining a levels of biomarkers RARRES2, LGALS3BP, and GPNMB in amniotic fluid; and (ii) comparing the levels determined in (i) to their levels in a control or to thresholds, wherein an increase in the level of the biomarker in the biological sample in comparison to their levels in the control or above the thresholds is indicative of presence of a symptomatic congenital cCMV infection in the fetus.
  • the threshold for RARRES2 is from 35 to 55 ng/ml and the threshold for LGALS3BP is from 2400 to 2500 ng/ml, as determined by the methods of the present invention.
  • the method further comprises diagnosing whether the subject, e.g. the fetus is infected with CMV
  • the method comprises measuring a viral load of CMV as known in the art in the biological sample, e.g. in an amniodc fluid.
  • the method further comprises a step of obtaining the biological sample.
  • the step of obtaining the biological sample is effected before determining the level of at least one biomarker.
  • the biological sample is obtained by any known method.
  • amniodc fluid is collected via amniocentesis.
  • the biological sample is further processed before determining the level of at least one biomarker, such as purification and dilutions.
  • the diagnosis is made before week 21 of pregnancy. According to some embodiments, the diagnosis is performed before week 24 of pregnancy. According to some embodiments, the diagnosis is made at week 12, 13, 14, 15, 16, 17, 18, 19 or week 20 of pregnancy. According to any one of the above embodiments, the diagnosis is made at or before week 36 of pregnancy.
  • the biological sample is obtained before week 21 of pregnancy. According to some embodiments, the biological sample is obtained before week 24 of pregnancy. According to some embodiments, the biological sample is obtained at week 12, 13, 14, 15, 16, 17, 18, 19 or week 20 of pregnancy. According to any one of the above embodiments, the biological sample is obtained at or before week 36 of pregnancy. According to other embodiments, the biological sample is obtained at week 12 to 36 of pregnancy. According to some embodiments, the biological sample is an amniotic fluid. Thus, according to some embodiments, the amniotic fluid is obtained at week 15, 16, 17 or 18 of the pregnancy. According to some embodiments, the amniotic fluid is obtained before week 24 of pregnancy. According to certain embodiments, the sample is fetal blood or serum.
  • the biological sample is maternal blood or serum.
  • the difference in the level of the at least one biomarker in the maternal blood or serum in comparison to the control is indicative of the presence of cCMV such as symptomatic cCMV infection in fetus.
  • the diagnosis is performed on or before week 36 of pregnancy
  • the diagnosis is made between week 12 to 36 of pregnancy.
  • the biomarker is a protein biomarker.
  • any known method may be used for the determination of the level of the at least one biomarker.
  • the biomarker is a protein biomarker and any method for determining the level of proteins or peptides may be used.
  • the method comprises use of an enzyme-linked immunosorbent assay (ELISA), luminex assay, or any other immune-detection assay.
  • the method comprises use of a mass spectroscopy.
  • the methods are chromatography methods.
  • an increase in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, and SCGB3A2 in the biological sample in comparison to its levels in the control is indicative of a symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of at least one biomarker of Group C and a decrease in the level of at least one biomarker of Group D in the biological sample in comparison to their levels in control is indicative of a symptomatic cCMV infection.
  • an increase in levels of 2, 3, 4 or 5 biomarkers selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, and SCGB3A2 and/or a decrease in levels of 2, 3, 4 or 5 biomarkers selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample in comparison to their levels in control is indicative of a symptomatic cCMV infection.
  • a biomarker selected from RARRES2, LGALS3BP and the combination thereof in the biological sample in comparison to their levels in control is indicative of a symptomatic cCMV infection.
  • the biological sample is amniotic fluid.
  • the amniotic fluid is obtained between weeks 12 to 36, or between weeks 14 to week 24 of pregnancy
  • the level of the biomarkers is determined by immunoassay method.
  • the biomarker is a protein biomarker.
  • the present invention provides a method of providing an indication useful in distinguishing between subjects infected with cCMV and uninfected, comprising (i) determining the level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the sample to its level in a control, wherein a difference in the levels of the biomarker in the biological sample and in the control provides an indication useful in distinguishing between infected and uninfected subjects, wherein at least one biomarker is selected from a biomarker of Group A and Group B.
  • the subject is a fetus.
  • the biological sample is amniotic fluid.
  • the present invention provides a method of providing an indication useful in distinguishing between subjects having a symptomatic cCMV and asymptomatic cCMV subjects, comprising (i) determining a level of at least one biomarker in a biological sample; and (ii) comparing the level of the at least one biomarker from the sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control provides an indication useful in distinguishing between symptomatic and asymptomatic cCMV or uninfected subjects, wherein at least one biomarker is selected from a biomarker of Group C and Group D.
  • the subject is a fetus.
  • the biological sample is amniotic fluid.
  • the present invention provides a method or diagnosing an asymptomatic cCMV infection in a subject, such as fetus.
  • a subject such as fetus.
  • the presence of infection in fetus may be carried out by the methods of the present invention or by any known method.
  • the fetus is infected but is not diagnosed as having a symptomatic CMV, said fetus is identified/diagnosed as having asymptomatic CMV.
  • a preventive treatment may be administered or adjusted.
  • the present invention provides a method of diagnosing an asymptomatic cCMV infection in a fetus comprising (i) diagnosing whether the fetus is infected with cCMV and (ii) diagnosing whether the fetus has a symptomatic cCMV according to the methods of the present invention.
  • a method for diagnosing whether the fetus is infected with cCMV can be used. Examples of such methods is as defined by the methods of the present invention or determining viral load e g.
  • the present invention provides a method of diagnosing an asymptomatic cCMV infection in fetus comprising (i) determining a level of at least one biomarker in a biological sample of an infected subject; and (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a lack of statistically significant difference in the level of the biomarker in the biological sample and in the control is indicative of presence of asymptomatic cCMV infection in the fetus, wherein the at least one biomarker is selected from the biomarkers of Group C and Group D.
  • the present invention provides a method of diagnosing an asymptomatic cCMV infection in the infected fetus comprising (i) determining a level of at least one biomarker in the biological sample; and (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a lack of statistically significant difference in the level of the biomarker in the biological sample and in the control is indicative of presence of asymptomatic cCMV infection in the fetus, wherein the at least one biomarker is selected from the biomarkers of Group C and Group D and the biological sample is an amniotic fluid.
  • the present invention provides a method for recommending a medical intervention, the method comprises detecting whether a subject has a congenital cytomegalovirus according to the methods of the present invention and recommending the medical intervention based on the obtained results. Any terms and definition according to the previous aspects and embodiment are valid and implemented in this aspect as well.
  • the present invention provides a method of diagnosing and recommending a medical intervention, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection; and (iii) providing a recommendation of a medical intervention based on the obtained results, wherein the biomarker is selected from CTSB, SIRPB1, LGALS3BP, AGRN, VCAM1, COTL1, CD5L, ATP5B, KRT6A, SECTM1, FABP3, S100A6, SH3BGRL, ISG15, CRTAC1, RARRES2, BST2, B2M, EPYC, DKK1, HLA-C, CPQ, F5, GPNMB, LAMAS, CA3, DES, MEGF8, GALNT7, FAT
  • the present invention provides a method of recommending a medical intervention, wherein the method comprises (i) determining a level of at least one biomarker in a biological sample; (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection; and (iii) providing a recommendation of a medical intervention based on the obtained results, wherein the biomarker is selected from CTSB, SIRPB1, LGALS3BP, AGRN, VCAM1, COTL1, CD5L, ATP5B, KRT6A, SECTM1, FABP3, S100A6, SH3BGRL, ISG15, CRTAC1, RARRES2, BST2, B2M, EPYC, DKK1, HLA-C, CPQ, F5, GPNMB, LAMAS, CA3, DES, MEGF8, GALNT7, FAT4, LPHN2, A
  • the cCMV is a symptomatic cCMV.
  • the present invention provides a method of recommending a medical intervention, wherein the method comprises 0) determining a level of at least one biomarker in a biological sample; fii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection; and (iii) providing a recommendation of a medical intervention based on the obtained results, wherein the biomarker is selected from biomarkers of Group C and Group D.
  • the cCMV is an asymptomatic CMV.
  • the present invention provides a method of recommending a medical intervention, wherein the method comprises (i) detecting whether the fetus has an asymptomatic CMV according to any one of the above aspects and embodiments and (ii) providing a recommendation of a medical intervention based on the obtained results.
  • the intervention comprises monitoring.
  • an increase in the level of at least one biomarker selected from SYNE1, MYH14, GPNMB, LG AL S3 BP, CPA2, RARRES2, and SCGB3A2 in the biological sample in comparison to its levels in the control is indicative of a symptomatic cCMV infection.
  • a decrease in the level of at least one biomarker selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample in comparison to their levels in the control is indicative of a symptomatic cCMV infection.
  • an increase in the level of at least one biomarker of Group C and decrease in the level of at least one biomarker of Group D in the biological sample in comparison to their levels in control is indicative of a symptomatic cCMV infection.
  • an increase in levels of 2, 3, 4 or 5 biomarkers selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, and SCGB3A2 and/or a decrease in levels of 2, 3, 4 or 5 biomarkers selected from CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, and GP1BA in the biological sample in comparison to their levels in control is indicative of a symptomatic cCMV infection.
  • an increase in levels of RARRES2 and LGALS3BP in the biological sample in comparison to their levels in control is indicative of a symptomatic cCMV infection.
  • the biological sample is amniotic fluid.
  • the amniotic fluid is obtained between week 12 to week 36 or between week 14 to week 24 of pregnancy.
  • the method further comprises a step of obtaining the biological sample.
  • the step of obtaining the biological sample is effected before determining the level of at least one biomarker.
  • the medical intervention is selected from a treatment of cCMV, cessation of treatment of cCMV, adjusting treatment, watchful waiting, monitoring or recommendation to terminate the pregnancy.
  • the treatment is pregnancy termination.
  • the term “adjusting treatment” contemplates cessation of treatment or changing the administered dose or changing the treatment.
  • the medical intervention comprises treatment and/or monitoring of a symptomatic cCMV.
  • the medical intervention comprises treatment and/or monitoring of an asymptomatic cCMV.
  • the present invention provides a method of providing a medical intervention in case of presence of symptomatic cCMV in fetus comprising detecting whether a fetus has a symptomatic cCMV by the methods of the present invention and treating the symptomatic cCMV, wherein medical intervention comprises treatment using an anti-CMV compound or pregnancy termination.
  • the present invention provides providing a medical intervention in case of presence of symptomatic cCMV in fetus comprising: (I) detecting whether a fetus has a symptomatic cCMV by (i) determining a level of at least one biomarker in a biological sample; (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in the control is indicative of a congenital CMV infection, and wherein the biomarker is selected from biomarkers of Group C and Group D; and (II) based on the result of (I) providing a medical intervention comprising treating cCMV or pregnancy termination.
  • the treatment of cCMV comprises treatment using an anti-CMV compound selected from an antiviral drug and antibodies against CMV
  • the antiviral drug is valacyclovir or letermovir.
  • the treatment comprises administering to the mother the anti-CMV compound.
  • the treatment comprises administering the fetus the anti-CMV compound.
  • the present invention provides a method of providing a medical intervention in case of presence of asymptomatic cCMV in fetus comprising detecting whether a fetus has an asymptomatic cCMV by the methods of the present invention and treating the symptomatic cCMV, wherein medical intervention comprises treatment using an anti-CMV compound.
  • the present invention provides providing a medical intervention in case of presence of asymptomatic cCMV in fetus comprising (I) determining whether the fetus in infected with
  • detecting whether a fetus has a symptomatic cCMV by (i) determining a level of at least one biomarker in a biological sample; (ii) comparing the level of the at least one biomarker from the biological sample to its level in a control, wherein a difference in the level of the biomarker in the biological sample and in die control is indicative of a congenital CMV infection, and wherein the biomarker is selected from biomarkers of Group C and Group D, and (III) based on the result of (I) and (II) providing a medical intervention comprising treating cCMV or cessation of treatment of the cCMV.
  • Determining whether the fetus is infected by CMV may be performed by the methods of the present invention or by any other methods such as determining the viral load of CMV by e.g. PCR.
  • the terms “treating”, “therapy” or “treatment” a condition or patient refers to taking steps to obtain beneficial or desired results, including clinical results.
  • Beneficial or desired clinical results include, but are not limited to, or ameliorating abrogating, substantially inhibiting, slowing or reversing the progression of a disease, condition or disorder, substantially ameliorating or alleviating clinical or esthetical symptoms of a condition, substantially preventing the appearance of clinical or esthetical symptoms of a disease, condition, or disorder, and protecting from harmful or annoying symptoms.
  • Treating further refers to accomplishing one or more of the following: (a) reducing the severity of the disorder, (b) limiting development of symptoms characteristic of the disorder(s) being treated; (c) limiting worsening of symptoms characteristic of the disorder(s) being treated; (d) limiting recurrence of the disorders) in patients that have previously had the disorders); and/or (e) limiting recurrence of symptoms in patients that were previously asymptomatic for the disorders).
  • administering or “administration of’ a substance, a compound or an agent to a subject can be carried out using one of a variety of methods known to those skilled in the art.
  • a compound or an agent can be administered intrauterinally, intra umbilical cord,intravenously, arterially, intradermally, intramuscularly, intraperitonealy, intravenously, subcutaneously, ocularly, sublingually, orally (by ingestion), intranasally (by inhalation), intraspinally, intracerebrally, and transdermally (by absorption, e g., through a skin duct).
  • a compound or agent can also appropriately be introduced by rechargeable or biodegradable polymeric devices or other devices, e g., patches and pumps, or formulations, which provide for the extended, slow or controlled release of the compound or agent.
  • Administering can also be performed, for example, once, a plurality of times, and/or over one or more extended periods.
  • the composition is administered 1 , 2, 3, 4, 5 or 6 times a day.
  • the composition is administered 1, 2, 3, 4, 5 or 6 times a month.
  • the administration includes both direct administration, including self-administration, and indirect administration, including the act of prescribing a drug.
  • a physician who instructs a patient to self-administer a drug, or to have the drug administered by another and/or who provides a patient with a prescription for a drug is administering the drug to the patient.
  • the present invention provide a method of treating a symptomatic or asymptomatic congenital cytomegalovirus in a fetus comprising diagnosing the symptomatic or asymptomatic cCMV in fetus according to the methods of the present invention and administering to the fetus or to the mother an anti-CMV active agent.
  • the present invention provides a method of determining the efficacy of the prevention or treatment of a symptomatic or asymptomatic congenital cytomegalovirus (cCMV) infection, the method comprises the steps of: (i) determining a level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group C and Group D; and (ii) comparing the level of the at least one biomarker from the biological sample to the level of the biomarker in a corresponding biological sample obtained from the same subject at an earlier sampling and/or to the level of the biomarker in the control, wherein a change in the level of the biomarkers in two consecutive measurements is indicative of efficacy of the prevention or therapy of the symptomatic or asymptomatic cCMV.
  • cCMV congenital cytomegalovirus
  • the present invention provides a method of determining efficacy of prevention of a congenital cytomegalovirus (cCMV) infection or disease, the method comprising the steps of: (i) determining the level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group A, Group B, Group C or Group D; and (ii) comparing the level of the at least one biomarker from the biological sample to the level of the biomarker in the control, wherein the efficacy of the prevention is evaluated according to the change or lack of change in the level(s) of the biomarker(s).
  • cCMV congenital cytomegalovirus
  • the present invention provides a method of determining the efficacy of therapy of a congenital cytomegalovirus (cCMV) infection or disease, the method comprising the steps of: (i) determining the level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group A, Group B, Group C or Group D; and (ii) comparing the level of the at least one biomarker from the biological sample to the level of the biomarker in a corresponding biological sample obtained from the same subject at an earlier sampling or to the level of the biomarker in the control, wherein the efficacy of the therapy is evaluated according to the change or lack of change in the level(s) of the biomarker(s).
  • cCMV congenital cytomegalovirus
  • the present invention provides a method of determining efficacy of prevention of a symptomatic cCMV infection or disease, the method comprising the steps of: (i) determining the level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group C or Group D; and (ii) comparing the level of the at least one biomarker from the biological sample to the level of the biomarker in the control or in a corresponding biological sample obtained from the same subject at an earlier sampling, wherein the efficacy of the prevention is evaluated according to the change or lack of change in the level(s) of the biomarker(s).
  • the present invention provides a method of determining the efficacy of therapy of a symptomatic cCMV, the method comprising the steps of: (i) determining the level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group C or Group D; and (ii) comparing the level of the at least one biomarker from the biological sample to the level of the biomarker in a corresponding biological sample obtained from the same subject at an earlier sampling or to the level of the biomarker in the control, wherein the efficacy of the therapy is evaluated according to the change or lack of change in the levd(s) of the biomarker(s) between subsequently obtained biological samples.
  • the present invention provides a method of determining efficacy of prevention of an asymptomatic cCMV infection or disease, the method comprising the steps of: (i) determining the level of at least one biomarker in a biological sample, wherein the at least one biomarker is selected from a biomarker of Group C or Group D; and (ii) comparing the level of the at least one biomarker from the biological sample to the level of the biomarker in a corresponding biological sample obtained from the same subject at an earlier sampling or to its level in the control, wherein the efficacy of the prevention is evaluated according to the change or lack of change in the level(s) of the biomarker(s).
  • the term “preventing” when used in relation to a condition refers to administration of a composition which reduces the frequency of, or delays the onset of, symptoms of the medical condition in a subject relative to a subject which does not receive the composition.
  • decreasing the difference between the level(s) of the biomarker(s) in the biological sample and its/their level in a control corresponds to effective treatment.
  • cessation of the increase of the difference between the level(s) of the biomarker(s) in the biological sample and its/their level in a control corresponds to effective treatment.
  • the present invention provides a kit comprising means for determining a level of at least one biomarker in a biological sample, and instructions for use, wherein the biomarker is selected from CTSB, SIRPB1, LGALS3BP, AGRN, VCAM1, COTL1, CD5L, ATP5B, KRT6A, SECTM1, FABP3, S100A6, SH3BGRL, ISG15, CRTAC1, RARRES2, BST2, B2M, EPYC, DKK1, HLA-C, CPQ, F5, GPNMB, LAMAS, CA3, DES, MEGF8, GALNT7, FAT4, LPHN2, AFM, MASP1, SOD3, GALNT2, LSR, HGF, PRG2, QSOX1, ERVMER34-1, PAPPA2, GOT1, RDX, IGFBP5, AHNAK, ADAM9, LAMP2, CPM, SBSN, CAST, SPINKS, TNMD, CDH15, KLKB1, PRSS
  • the kit comprises means for determining a level of at least one biomarker in a biological sample, and instructions for use, wherein the biomarker is selected from SYNE1, MYH14, GPNMB, LGALS3BP, CPA2, RARRES2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS1, PPBP, GP1BA and any combination thereof.
  • the kit comprises means for comparing the level of the biomarker in the biological sample to its level in a control.
  • the kit comprises means for comparing the level of the biomarker in the biological sample to its level in another biological sample.
  • the kit comprises means for comparing the level of the biomarker in the biological sample to a predefined threshold.
  • the kit is an assay kit.
  • the kit is a diagnostic kit.
  • the instructions comprise instructions for diagnosing a congenital cytomegalovirus (cCMV) infection.
  • the instructions comprise instructions for diagnosing a symptomatic congenital cytomegalovirus (cCMV) infection.
  • the instructions comprise levels of the at least one biomarker in the control.
  • the instructions comprise instructions of comparing the levels in the biological sample to the levels in the control or in another biological sample.
  • the instructions comprise instructions of comparing the levels of the biomarkers in the biological sample to a predefined threshold.
  • the kit comprises means for determining the levels of the at least one biomarker in the control or in another biological sample.
  • the instructions comprise means to differentiate between symptomatic and asymptomatic infected subjects.
  • the cCMV is a symptomatic cCMV.
  • the means for determining the level of at least one biomarker comprises an agent specific for determining the level of the at least one biomarker.
  • the agent is an antibody capable of binding to the at least one biomarker or a combination of antibodies.
  • the antibodies are selected from monoclonal antibodies, secondary antibody capable of binding an antibody specific to the at least one biomarker, tagged antibody such as fluorescently tagged, and any combination thereof.
  • the means for determining the level of at least one biomarker in a biological sample is an immunoassay kit.
  • the means is ELISA kit.
  • the means are means for performing ELISA.
  • the means are means for performing immunoassay test.
  • the present invention provides a kit comprising at least one ELISA kit for determining the level of at least one biomarker in a biological sample, and instructions for use, wherein the biomarker is selected from a biomarker of Group A, Group B, Group C and Group D.
  • the present invention provides a kit comprising at least one ELISA kit for determining the level of at least one biomarker in a biological sample, and instructions for use, wherein the biomarker is selected from a biomarker of Group C and Group D.
  • the means is any method capable of determining levels of a plurality of biomarkers, such as luminex.
  • the present invention provides a kit comprising means for determining a level of at least one biomarker from biomarkers of Group C and D in a biological sample, and instructions for use in diagnosis asymptomatic congenital CMV infection wherein the instructions to diagnose asymptomatic CMV comprise instructions to detect whether the subject is infected with CMV and negating that the subject has a symptomatic CMV.
  • the kit of the present invention comprises means for determining the level of RARRES2 in the biological sample. According to other embodiments, the kit of the present invention comprises means for determining the level of LGALS3BP. According to one embodiment, the kit of the present invention comprises means for determining the levels of RARRES2 and LGALS3BP. According to some embodiments, the kit comprises means for determining the level of GPNMB. According to some embodiments, the kit comprises means for determining the levels of RARRES2 and GPNMB or LGALS3BP and GPNMB. According to one embodiment, the kit of the present invention comprises means for determining the levels of RARRES2, LGALS3BP and GPNMB.
  • the kit comprises means for determining the level of SYNE1. According to some embodiments, the kit comprises means for determining the level of ANGPTL6. According to some embodiments, the kit comprises means for determining the level of GAS1. According to some embodiments, the kit comprises means for determining the level of CHGB. According to some embodiments, the kit comprises means for determining the level of ADAM9. According to some embodiments, the kit of the present invention further comprises means for determining a level of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, ADAM9, GAS 1 , PPBP, and GP1BA.
  • the kit of the present invention comprises means for determining levels of RARRES2 and LG AL S3 BP and further comprises means for determining a level of at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, SCGB3A2, CPM, PDGFRB, ANGPTL6, DMKN, CHGB, AD AM9, GAS 1 , PPBP, and GP 1 BA.
  • the means is ELISA kit.
  • the present invention provides a kit comprising at least one ELISA kit for determining the level of at least one biomarker in a biological sample, and instructions for use, wherein the biomarker is selected from RARRES2, LGALS3BP and a combination thereof.
  • the kit of the present invention further comprises ELISA kit for determining the level at least one biomarker selected from SYNE1, MYH14, GPNMB, CPA2, SCGB3A2 in the biological sample.
  • the kit comprises any other means to determine the levels of a plurality of biomarkers, instead of ELISA kit.
  • the kit further comprises reference levels of the biomarkers as in the control or means to determine the reference levels of the biomarkers or numerical values of the levels of the biomarker in the control or the threshold value of the biomarker above or below which the subject is diagnosed as having a cCMV and/or having a symptomatic cCMV.
  • the terms “reference level” and “threshold” are used herein interchangeably and refer to a level of the biomarker in the control above which or below which or in comparison to which a subject is diagnosed according to the teaching of the present invention, i.e. having cCMV infection and/or having a symptomatic cCMV infection.
  • the kit further comprises means for processing the biological sample.
  • the processing the biological sample comprises any procedure to allow measurement or improve measurement of the levels of the biomarkers, such as purification, dilution, concentration, separation of biomarkers etc.
  • the kit is for use in diagnosing a congenital CMV infection.
  • the cCMV is a symptomatic CMV, thus the kit is for use in diagnosing or detecting a symptomatic congenital CMV infection.
  • the cCMV is an asymptomatic CMV, thus the kit is for use in diagnosing or detecting an asymptomatic congenital CMV infection.
  • the present invention provides a use of a kit comprising means for determining the level of at least one biomarker in a biological sample, in the diagnosis of a congenital CMV, wherein the at least one biomarker is selected from a biomarker of Group A, Group B, Group C and Group D.
  • the cCMV is a symptomatic cCMV.
  • the at least one biomarker is selected from a biomarker of Group C and Group D.
  • the instructions comprise means to differentiate between symptomatic and asymptomatic infected subjects.
  • the present invention provides a use of a kit comprising means for determining the level of at least one biomarker in a biological sample, in diagnosis of a symptomatic congenital CMV, wherein the at least one biomarker is selected from a biomarker of Group C and Group D.
  • Amniotic fluid samples were obtained from women with primary CMV infection at 21-23 weeks' gestation as part of the routine diagnosis of fetal infection.
  • peptides were analyzed using nanoflow liquid chromatography (nanoAcquity) coupled to high resolution, high mass accuracy mass spectrometry (Q Exactive HFX). Each sample was analyzed on the instrument separately in a random order in discovery mode.
  • Raw data was processed with Max Quant vl .6.0.16.
  • the data was searched with the Andromeda search engine against the human and HCMV proteome databases appended with common lab protein contaminants and the following modifications: Caibamidomethylation of C as a fixed modification and oxidation of M, deamidation of N and Q and protein N- terminal acetylation as variable ones.
  • LGALS3BP Galectin-3-binding protein
  • the combined assay results (performed for a total of 20 symptomatic and 29 asymptomatic cases) confirmed the significant differences between the concentrations of LG AL S3 BP in amniotic fluid samples of symptomatic versus asymptomatic cCMV cases (p ⁇ 0.001; Mann-Whitney test and student T-test), and allowed for the definition of cutoff concentration values for reliable prenatal disease prediction, (for example, sensitivity of 90.5%, specificity of 96.4%, positive predictive value of 95.0%, and negative predictive value of 93.1% for a cutoff concentration of 2475 ng/ml).

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