Rapid diagnosis of peritonitis in peritoneal dialysis patients
Described is an assay for diagnosing an infection such as peritonitis in a subject. The assay includes a binding molecule, such as an antibody that specifically binds to an inflammatory marker in a sample from the subject, and a second binding molecule that binds to a marker indicative of a pathogen in the sample. For diagnosing peritonitis in a subject, the pathogen will be at least one bacterium and/or fungus. Typically, the assay will be incorporated into a lateral flow device and may include a binding molecule that specifically binds to an antigen indicative of the presence of a specific pathogen species. The described assay(s) may further include filter(s), enriching antigen(s), and buffer(s). Also described are methods of diagnosing and treating peritonitis in a subject who is a peritoneal dialysis patient that include utilizing the herein described assay(s) or assay kit(s) to analyze the subject's peritoneal dialysis effluent.
1. An assay kit for analyzing a sample from a subject, the assay kit comprising:
a binding molecule that specifically binds an antigen indicative of a subject launching an inflammatory response in the peritoneum, wherein the antigen indicative of a subject launching an inflammatory response in the peritoneum is neutrophil gelatinase-associated lipocalin (NGAL);
a binding molecule that specifically binds an antigen indicative of the presence of Gram-negative bacteria, wherein the antigen indicative of the presence of Gram-negative bacteria is lipopolysaccharide (LPS);
a binding molecule that specifically binds an antigen indicative of the presence of Gram-positive bacteria, wherein the antigen indicative of the presence of Gram-positive bacteria is lipoteichoic acid (LTA); and
a binding molecule that specifically binds an antigen indicative of the presence of Staphylococcus aureus , wherein the an antigen indicative of the presence of S. aureus is peptidoglycan (PGN).
2. The assay kit of claim 1 , further comprising:
a binding molecule that specifically binds an antigen indicative of the presence of a fungus, wherein the antigen indicative of the presence of a fungus is β-glucan.
3. An assay kit for analyzing a sample from a subject, the assay kit comprising:
a binding molecule that specifically binds an antigen indicative of a subject launching an inflammatory response in the peritoneum, wherein the antigen indicative of a subject launching an inflammatory response in the peritoneum is neutrophil gelatinase-associated lipocalin (NGAL);
a binding molecule that specifically binds an antigen indicative of the presence of Gram-negative bacteria, wherein the antigen indicative of the presence of Gram-negative bacteria is lipopolysaccharide (LPS);
a binding molecule that specifically binds an antigen indicative of the presence of Gram-positive bacteria, wherein the antigen indicative of the presence of Gram-positive bacteria is lipoteichoic acid (LTA); and
a binding molecule that specifically binds an antigen indicative of the presence of a fungus.
4. The assay kit of claim 3 , wherein β-glucan is the antigen indicative of the presence of a fungus.
5. The assay kit of claim 3 , further comprising:
a binding molecule that specifically binds an antigen indicative of the presence of a specific pathogen species in a peritoneal dialysis effluent.
6. The assay kit of claim 5 , wherein the specific pathogen is selected from the group consisting of Staphylococcus aureus , Methicillin-resistant Staphylococcus aureus , a Coagulase-negative staphylococcal species Staphylococcus epidermidis, Staphylococcus haemolyticus , a Streptococcus sp., an Escherichia coli , a Candida sp., an Aspergillus sp., and a vancomycin-resistant Enterococcus sp.
7. The assay kit of claim 3 , further comprising:
means for filtering, concentrating and/or enriching antigen(s) from peritoneal dialysis effluent.
8. The assay kit of claim 3 , further comprising:
a buffer that elutes antigen(s) from a sample from a subject.
9. A method of diagnosing peritonitis in a peritoneal dialysis patient, the method comprising:
contacting peritoneal dialysis effluent of the patient with the binding molecules of the assay kit of claim 3 so as to diagnose peritonitis by determining whether the patient is launching an inflammatory response in the peritoneum, whether Gram-negative bacteria are present in the peritoneal dialysis effluent, whether Gram-positive bacteria are present in the peritoneal dialysis effluent, and/or whether fungus is present in the peritoneal dialysis effluent.
10. A method of treating peritonitis in a peritoneal dialysis patient, the method comprising:
contacting peritoneal dialysis effluent of the patient with the binding molecules of the assay kit of claim 3 so as to diagnose peritonitis by determining whether the patient is launching an inflammatory response in the peritoneum, whether Gram-negative bacteria are present in the peritoneal dialysis effluent, whether Gram-positive bacteria are present in the peritoneal dialysis effluent, and/or whether fungus is present in the peritoneal dialysis effluent, and then
administering an appropriate antibiotic to the patient to treat the peritonitis in view of the diagnosis.
11. A method of diagnosing peritonitis in a peritoneal dialysis patient, the method comprising:
contacting peritoneal dialysis effluent of the patient with the binding molecules of the assay kit of claim 3 , and
using the binding molecule that specifically binds NGAL to detect an antigen indicative of the patient having launched an inflammatory response in the peritoneum that may be present in the peritoneal dialysis effluent;
using the binding molecule that specifically binds LTA to detect in the peritoneal dialysis effluent an antigen indicative of the presence of Gram-positive bacteria that may be present in the peritoneal dialysis effluent;
using the binding molecule that specifically binds LPS to detect in the peritoneal dialysis effluent an antigen indicative of the presence of Gram-negative bacteria that may be present in the peritoneal dialysis effluent, and
using the binding molecule that specifically binds an antigen indicative of the presence of a fungus to detect in the peritoneal dialysis effluent an antigen indicative of the presence of a fungus,
so as to diagnose peritonitis by determining whether the patient is launching an inflammatory response in the peritoneum, whether Gram-negative bacteria are present in the peritoneal dialysis effluent, whether Gram-positive bacteria are present in the peritoneal dialysis effluent, and/or whether fungus is present in the peritoneal dialysis effluent.
12. The method according to claim 11 , wherein β-glucan is the antigen indicative of the presence of a fungus.
13. The method according to claim 11 , further comprising:
detecting in the peritoneal dialysis effluent an antigen indicative of the presence of a specific pathogen species in a peritoneal dialysis effluent.
14. The method according to claim 13 , wherein the specific pathogen is selected from the group consisting of Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, Escherichia coli , a Candida sp., and a vancomycin-resistant Enterococcus sp.
15. The method according to claim 11 , further comprising:
filtering, concentrating, and/or enriching the peritoneal dialysis effluent prior to detecting antigen(s).
16. The assay kit of claim 3 , further comprising:
a binding molecule that specifically binds peptidoglycan (PGN).