Methods for detecting bordetella

A genus and detection marker technology, applied in the field of detection of Bordetella, can solve problems such as false negative results

Pending Publication Date: 2019-01-04
QUEST DIAGNOSTICS INVESTMENTS INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These assays may give false negative results if patient samples are not obtained within the appropriate time window during disease progression

Method used

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  • Methods for detecting bordetella
  • Methods for detecting bordetella
  • Methods for detecting bordetella

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0166] Example 1: Detection of Pathogenic Bordetella Species Using SIMPLEXA Direct Real-Time PCR Nasopharyngeal swab samples were collected from patients. 50 μL of raw and undiluted nasal swab samples were loaded directly into sample wells of wedge 1 of SIMPLEXA direct amplification discs (Focus Diagnostics, Inc., Cypress, CA, USA) without a separate front-end sample preparation step .

[0167] Aspirate 50 μL of the amplification master mix into the reaction well of wedge 1 of the plate, wherein the amplification master mix includes PCR buffer, DNA polymerase, dNTPs, magnesium chloride, potassium chloride, prepared by SEQ ID NO:4 -primers consisting of 5, 7-8 and 10-11, probes consisting of SEQ ID NO: 6, 9 and 12 and an internal control DNA fragment (SEQ ID NO: 13) and a pair of primers specific for the control fragment and Probes (SEQ ID NO: 17-19). TaqMan probes consisting of SEQ ID NO: 6, 12, 9 and 19 were labeled with JOE, FAM, CFR610 and Q670, respectively. Positive ...

Embodiment 2

[0175] Example 2: Limits of Detection of Bordetella Multiplex Assay

[0176] Limit of detection (LoD) studies were performed to determine the Simplexa TM Analytical Sensitivity of Bordetella Direct Assay in Nasopharyngeal Swab Matrix. Each bacterial strain will be screened for putative LoD and subsequently confirmed. The LoD for each strain was determined as the lowest concentration with >85-95% detection (>19 out of 20 replicates) for 20 replicates.

Embodiment 3

[0177] Example 3: Cross-reactivity of Bordetella multiplex assays

[0178] For cross-reactivity determinations, control nasal swab samples will be spiked with one of the test organisms listed below (n=5 for each organism).

[0179] Bacillus cereus

Chlamydophila pneumoniae

Haemophilus influenzae

Klebsiella pneumonia

Legionella pneumophila

Mycoplasma pneumoniae

Streptococcus pneumoniae

Staphylococcus aureus

Moraxella catarrhalis

Influenza A virus (Influenza A)

Influenza B virus

RSVB

[0180] A Bordetella multiplex assay was performed on each sample. In each case, a Ct value ≤40 was interpreted as a positive result for B. pertussis cross-reactivity, a Ct value ≤40 was interpreted as a positive result for B. parapertussis cross-reactivity, and a Ct value of ≤40 was interpreted as a positive result for B. hallii cross-reactivity.

[0181] No cross-reactivity is expected to be observed for any of the above ...

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Abstract

The present disclosure provides methods for determining whether a patient exhibiting pertussis-like symptoms will benefit from treatment, with therapeutic agents that inhibit Bordetella holmesii. These methods are based on detecting Bordetella pertussis, Bordetella parapertussis, and Bordetella holmesii in a biological sample by assaying for the presence of the IS481, IS 1001, and hIS1001 target repeat elements, respectively. Kits for use in practicing the methods are also provided.

Description

[0001] Cross References to Related Applications [0002] This application claims the benefit of and priority to US Application No. 62 / 312,883, filed March 24, 2016, the contents of which are hereby incorporated by reference in their entirety. technical field [0003] The present disclosure provides methods for determining whether a patient exhibiting pertussis-like symptoms would benefit from treatment with a therapeutic agent that inhibits Bordetella holmesii. These methods are based on the detection of Bordetella pertussis, Bordetella parapertussis and Bordetella hockii in biological samples by determining the presence of IS481, IS1001 and hIS1001 targeted repeats. bacteria. Kits for carrying out the methods are also provided. Background technique [0004] The following description of the background of the present disclosure is provided only to assist the reader in understanding the present disclosure and is not admitted to describe or constitute prior art to the present...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07H21/04C07H21/00
CPCC12Q1/689C12Q2600/158C12Q2600/16A61K31/407A61K31/496A61K31/7048C12Q2600/106
Inventor J·陈M·泰伯
Owner QUEST DIAGNOSTICS INVESTMENTS INC
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