Detection of intraamniotic infection

• US 9,164,092 B2
• Filed: 02/07/2014
• Issued: 10/20/2015
• Est. Priority Date: 11/25/2009
• Status: Active Grant

• Current Assignee: Hologic Incorporated
• Original Assignee: Hologic Incorporated
• Inventors: Grove, Thomas H.
• Primary Examiner(s): COOK, LISA V
• Application Number:
  US14/175,476
• Publication Number:
  US 20140154714A1
• Time in Patent Office: 620 Days
• Field of Search: 435/7.1, 435/7.21, 436/501, 436/518, 424/9.1, 424/520, 422/430, 530/300, 530/350
• US Class Current: 1/1
• CPC Class Codes: G01N 2333/47 Assays involving proteins of known structure or function as defined in the subgroups
• CPC Class Codes: G01N 2333/5412 IL-6
• CPC Class Codes: G01N 2800/36 Gynecology or obstetrics
• CPC Class Codes: G01N 2800/368 Pregnancy complicated by disease or abnormalities of pregnancy, e.g. preeclampsia, preterm labour
• CPC Class Codes: G01N 33/56933 Mycoplasma
• CPC Class Codes: G01N 33/6869 Interleukin
• CPC Class Codes: G01N 33/689 related to pregnancy or the gonads

Abstract

The present invention concerns the identification of biomarkers and groups or combinations of biomarkers that can be used for non-invasive diagnosis of intra-amniotic infection, and diagnostic assays using such biomarkers.

Claims

1. A method for the diagnosis of intra-amniotic inflammation or infection in a pregnant female mammalian subject comprising:

(a) measuring in a sample of cervical-vaginal fluid obtained from said subject the levels of interleukin-6 (IL-6) and fetal fibronectin (fFN) relative to the corresponding levels of said proteins in normal cervical-vaginal fluid or cervical-vaginal fluid known to be indicative of intra-amniotic inflammation or infection; and

(b) diagnosing said subject with intra-amniotic inflammation or infection if each of said levels of each of said proteins in said sample is determined to show a statistically significant difference relative to the corresponding levels of each of said proteins in said normal cervical-vaginal fluid, or is determined not to show a statistically significant difference relative to the corresponding levels of each of said proteins in said cervical-vaginal fluid known to be indicative of intra-amniotic inflammation or infection.

2. The method of claim 1 wherein the subject is a human patient.

3. The method of claim 1 wherein said protein level is determined by methods comprising the use of an immunoassay, a protein array, an immunochromatographic test, mass spectrometry, or combinations thereof.

4. The method of claim 3 wherein said level is determined using an immunochromtaographic test employing a lateral flow device.

5. The method of claim 4, wherein the lateral flow device is an immunochromatographic strip (ICS) test.

6. The method of claim 3 wherein said immunoassay employs antibodies and reagents for the detection of IL-6 and fFN.

7. The method of claim 6, wherein the antibodies and reagents comprise a capture antibody and a detector antibody.

8. The method of claim 7, wherein the capture antibody and detector antibody are monoclonal antibodies.

9. The method of claim 7, wherein the capture antibody and detector antibody are polyclonal antibodies.

10. The method of claim 7, wherein the capture antibody and detector antibody are either monoclonal or polyclonal antibodies.

11. The method of claim 8, wherein the antibodies comprise monoclonal antibodies having binding specificity for IL-6 and fFN, and wherein the method further comprises use of an anti-antibody immunoglobulin.

12. The method of claim 11, wherein the monoclonal antibodies and the anti-antibody immunoglobulin are provided in an amount of about 0.001 mg to about 100 grams.

13. The method of claim 11, wherein the monoclonal antibodies and the anti-antibody immunoglobulin are provided in an amount of about 0.01 mg to about 1 gram.

14. The method of claim 11, wherein the anti-antibody immunoglobulin is selected from the group consisting of a polyclonal immunoglobulin, protein A and protein G, or functional fragments thereof.

15. The method of claim 14, wherein the anti-antibody immunoglobulin is labeled.

16. The method of claim 6, further comprising use of agents for reducing background interference in a test or agents for increasing signal.

17. The method of claim 6, further comprising use of software and algorithms for combining and interpolating marker values to produce a prediction of clinical outcome of interest.

18. The method of claim 6, further comprising use of an apparatus for conducting a test.

19. A method for determining signs and symptoms indicating intra-amniotic inflammation or infection comprising

(a) measuring in a sample of cervical-vaginal fluid obtained from said subject the levels of interleukin-6 (IL-6) and fetal fibronectin (fFN), relative to the levels of IL-6 and fFN in normal cervical-vaginal fluid or cervical-vaginal fluid known to be indicative of intra-amniotic inflammation or infection;

and

(b) diagnosing said subject with intra-amniotic inflammation or infection if each of said levels of IL-6 and fFN in said sample are determined to show a statistically significant difference relative to the corresponding levels of IL-6 and fFN in said normal cervical-vaginal fluid, or are determined not to show a statistically significant difference relative to the corresponding levels of IL-6 and fFN in said cervical-vaginal fluid known to be indicative of intra-amniotic inflammation or infection.

20. The method of claim 19, wherein the signs and symptoms comprise maternal fever, maternal leukocytosis, maternal and/or fetal tachycardia, uterine tenderness, and/or foul-smelling amniotic fluid.

21. The method of claim 19 wherein said protein level is determined by methods comprising the use of an immunoassay, a protein array, an immunochromatographic test, mass spectrometry, or combinations thereof.

22. The method of claim 20, wherein the maternal fever is >

37.8°

C.

23. The method of claim 20, wherein the maternal leukocytosis is >

15,000 /mm³ .

24. The method of claim 19, wherein said protein level is determined by methods comprising the use of an immunoassay, a protein array, an immunochromatographic test, mass spectrometry, or combinations thereof.