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Methods for screening infections

A technology for trypanosoma cruzi infection and subjects, applied in the field of screening infection, which can solve the problems of intensive and complicated labor, and the inability of diagnostic tools to reliably detect the early stage of infection

Pending Publication Date: 2019-12-06
COWPER SCI INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Current methods are time-consuming, complex, and labor-intensive, and may require varying degrees of expertise
Additionally, available diagnostic tools are often not reliable for detecting early stages of infection, and more than one method is often required to positively diagnose infection

Method used

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  • Methods for screening infections
  • Methods for screening infections
  • Methods for screening infections

Examples

Experimental program
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Effect test

Embodiment 4

[0123] Example 4 illustrates a method for identifying candidate protein biomarkers using a discriminative peptide that differentiates serological status in samples from healthy subjects from those from subjects infected with Trypanosoma cruzi (Chagas disease). sample. A healthy subject may be a subject who was previously infected with Trypanosoma cruzi and seroconverted to seronegative, and / or a subject who has never been infected with Trypanosoma cruzi. Table 2 provides a list of candidate protein biomarkers. Similarly, candidate protein biomarkers can be identified using discriminative peptides that distinguish samples from subjects with other infectious diseases from samples from healthy subjects, samples from subjects with other infectious diseases and the serological status of samples from subjects with simulated disease (which may or may not be infectious).

[0124] In some embodiments, a method for identifying a candidate protein biomarker for an infectious disease co...

Embodiment 1

[0196] Example 1 - Immunosignature method for diagnosing infection

[0197] An immunosignature assay was developed to detect and differentiate T. cruzi, HBV, HCV, and WNV infections according to the following.

[0198] Donor samples. Plasma samples from donors seropositive for Chagas antibodies, as well as age- and sex-matched healthy donor plasma, and for hepatitis B virus (HBV), hepatitis C virus (HCV), or West Nile virus (WNV) Plasma samples that tested seropositive were obtained from Creative Testing Solutions (Tempe, AZ). Two sample cohorts were obtained, one in 2015 and the other in 2016. Upon receipt, plasma was thawed, mixed 1:1 with ethylene glycol as a cryoprotectant, and aliquoted into single-use volumes. Single-use aliquots were stored at -20°C until needed. The remaining sample volume was stored neat at -80 °C. The identity of all samples was tracked using tubes (Micronic, Leystad, the Netherlands) with two-dimensional barcodes. In preparation for the assay,...

Embodiment 2

[0220] Example 2 - Platform Verification

[0221] Experiments were performed using monoclonal antibodies to evaluate the quality of the final in situ synthesized array peptide products with respect to ligand presentation and antibody recognition.

[0222] All diagnostic assays are performed on a validated microarray platform.

[0223] A protocol for peptide synthesis was developed in which parallel coupled reactions were performed directly on silicon wafers using masking and photolithography. Antibody binding events were queried using arrays representing a total of 131,712 peptides (median length 9 amino acids) at each 14 μm x 14 μm feature. The array layout included 126,009 library peptide features and 6203 control peptide features attached to the surface via a common linker (see Example 1). The library peptides were designed to sample all possible amino acid combinations evenly. Control peptides included 500 features corresponding to established epitopes of five different...

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Abstract

The disclosed embodiments concern non-invasive methods, and apparatus, and systems for identifying infections. The methods are predicated on identifying discriminating peptides present on a peptide array, which are differentially bound by the different mixtures of antibodies present in samples from subjects consequent to an infection relative to binding of mixtures of antibodies present in reference subjects.

Description

[0001] cross reference [0002] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 462,320, filed February 22, 2017, which is hereby incorporated by reference in its entirety. Background technique [0003] Infectious diseases are conditions usually caused by microorganisms such as bacteria, viruses, fungi or parasites. Diagnosis of infection usually requires laboratory tests of bodily fluids such as blood, urine, throat swabs, stool samples and, in some cases, spinal fluid. Imaging scans and biopsies can also be used to identify the source of infection. A variety of individual tests are available for diagnosing infection, and include immunoassays for the pathogen, polymerase chain reaction, fluorescent in situ hybridization, and genetic testing. Current methods are time-consuming, complex, and labor-intensive, and may require varying degrees of expertise. Additionally, available diagnostic tools often cannot reliably detect early stages of in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K9/00C07K14/47C40B30/04C40B40/10G01N33/564
CPCG01N33/56905G01N33/5761G01N33/5767G01N2333/183G01N2469/20G01N33/68G01N2333/44G01N2570/00C12Q1/70
Inventor 凯瑟琳·弗朗西丝·赛克斯罗伯特·威廉·葛文乔纳森·斯科特·梅尔尼克迈克尔·威廉·罗西奥多·迈克尔·塔拉索
Owner COWPER SCI INC
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