Streptococcus pneumoniae serotype specific pcr pad assay
AssigneeMerck Sharp & Dohme Corp.
InventorsKate L. Hicks, Gowrisankar Rajam, Leonard J. Rubinstein
Life of this patent
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Abstract
The invention described herein relates to a high-throughput and multiplex Streptococcus pneumoniae (S. pneumoniae) serotype (ST)-specific polymerase chain reaction-pneumococcal antigen detection (PCR-PAD) assay useful to support epidemiology studies, clinical trials and clinical therapy. The invention further provides kits for detecting the presence or absence of S. pneumoniae and the presence or absence of particular S. pneumoniae strains in a patient sample.
Claims13
An assay for detecting S. pneumoniae pneumococcal serotypes (STs) in a patient sample comprising i) using direct polymerase chain reaction (PCR) to detect the presence of highly conserved S. pneumoniae nucleic acid sequences in the sample indicating a sample that contains S. pneumoniae (a positive sample);
- and ii) screening the positive sample to identify one or more particular pneumococcal polysaccharide (PnPs) STs present in the positive sample comprising:
- a) contacting the positive sample with:
- one or more first monoclonal antibodies that can bind one or more particular PnPs STs in the sample to form one or more first mAb-antigen complexes;
- b) contacting the positive sample with:
- one or more second monoclonal antibodies that can bind the same one or more particular PnPs STs to form one or more first mAb-antigen-second mAb complexes;
- c) contacting the positive sample with:
- a third antibody that can bind the one or more second monoclonal antibodies, wherein the third antibody is detectably coupled to a reporter molecule;
- d) detecting the presence or absence of the one or more first mAb-antigen-second mAb complexes by detecting the reporter molecule;
- wherein the presence of the one or more first mAb-antigen-second mAb complexes confirms the presence of particular PnPs STs and therefore the presence of particular STs of S. pneumoniae;
- and
- wherein the absence of the one or more first mAb-antigen-second mAb complexes is indicative of the absence of particular PnPs STs and therefore the absence of particular STs of S. pneumoniae.
A kit for detecting the presence or absence of S. pneumoniae and the presence or absence of particular STs of S. pneumoniae in a sample, wherein said kit comprises:
- a) one or more PCR primers that bind highly conserved S. pneumoniae nucleic acid sequences;
- b) one or more first monoclonal antibodies that bind one or more pneumococcal capsular PnPs STs;
- c) one or more second monoclonal antibodies that bind one or more PnPs STs;
- d) a third antibody that binds the one or more second monoclonal antibodies;
- and
- e) instructions to use said kit.
Dependencies are inferred from the claim text.