Selective detection of <i>Bordetella </i>species

摘要

A process for detecting Bordetella spp. nucleic acid in a biological sample includes producing an amplification product(s) by amplifying one or more Bordetella spp. in a multiplex single chamber PCR assay, and measuring the amplification product(s) to detect or distinguish Bordetella spp. in the biological sample. Also provided are reagents and methods for detecting and distinguishing Bordetella spp. from each other and other bacteria or viruses. A kit is provided for detecting and quantifying one or more Bordetella spp. in a biological sample.

主权项

1. A process of detecting and distinguishing Bordetella pertussis, B. parapertussis, and B. holmesii in a biological sample comprising:
producing a first amplification product by amplifying a Bordetella nucleotide sequence if present in the sample using a forward primer consisting of the sequence of SEQ ID NO: 1 and a reverse primer consisting of the sequence of SEQ ID NO: 2, under conditions suitable for a polymerase chain reaction; measuring said first amplification product; producing a second amplification product by amplifying a Bordetella nucleotide sequence if present in the sample using a forward primer consisting of the sequence of SEQ ID NO: 4 and a reverse primer consisting of the sequence of SEQ ID NO: 5, under conditions suitable for a polymerase chain reaction, and measuring said second amplification product; producing a third amplification product by amplifying a Bordetella nucleotide sequence if present in the sample using a forward primer consisting of the sequence of SEQ ID NO: 7 and a reverse primer consisting of the sequence of SEQ ID NO: 8, under conditions suitable for a polymerase chain reaction, and measuring said third amplification product; said step of producing said first amplification product, said step of producing said second amplification product, and said step of producing said third amplification product performed in the same reaction chamber; producing a fourth amplification product by amplifying a Bordetella nucleotide sequence using a forward primer consisting of the sequence of SEQ ID NO: 10 and a reverse primer consisting of the sequence of SEQ ID NO: 11, under conditions suitable for a polymerase chain reaction, and measuring said fourth amplification product; detecting and distinguishing Bordetella pertussis, B Bordetella parapertussis, and B Bordetella holmesii in the sample based on a relative ratio of said first amplification product, said second amplification product, said third amplification product, and said fourth amplification product.