Compositions and methods for detection of Mycoplasma genitalium
Inventors
Harris, Jody • Mangipudi, Kalyani
Assignees
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Publication Number
US-12460267-B2
Publication Date
2025-11-04
Expiration Date
Abstract
Methods for the rapid detection of the presence or absence of Mycoplasma genitalium (MG) in a biological or non-biological sample are described. The methods can include performing an amplifying step, a hybridizing step, and a detecting step. Furthermore, primers, probes targeting the target MG gene, along with kits are provided that are designed for the detection of MG.
Core Innovation
The invention relates to rapid nucleic-acid diagnostic methods and kits for detecting Mycoplasma genitalium (MG). The disclosed approach uses cycling-based amplification together with hybridization and fluorescent detection, including real-time PCR with fluorescent hydrolysis probes and fluorescence donor/acceptor formats such as FRET. The kit is configured so that a fluorescent detectably labeled probe hybridizes to an amplicon generated by a forward primer and a reverse primer.
Detection is based on MG nucleic acid targets including 23S ribosomal RNA (23S rRNA), the conserved region A of the mgpB gene, and multi-copy MgPar partial repeats (EF region). Specific forward primer, reverse primer, and fluorescent detectably labeled probe oligonucleotide sequences are selected from enumerated SEQ ID NO sets, and the probe is configured to hybridize to the MG-generated amplicon. The disclosure also includes sequence-labeling formats using fluorescent donor and fluorescent acceptor moieties and quencher labeling.
The disclosed diagnostic design addresses specificity with respect to Mycoplasma pneumoniae and optionally supports multiplexing with Trichomonas vaginalis (TV). The disclosure further describes optional confirmation using dye-binding and melting temperature or melting-curve analysis. Example results include reported real-time PCR performance and limit of detection values, including MG limit of detection below 1 genomic equivalent per PCR in co-amplification with an internal control standard.
Claims Coverage
The partial claim set provides one independent claim directed to a kit for detecting MG nucleic acid using specifically selected forward and reverse primers and a fluorescently detectably labeled probe, where the probe hybridizes to an MG-generated amplicon. The independent-claim inventive content is implemented through defined SEQ ID NO selection ranges for each oligonucleotide component.
Fluorescently labeled probe configured to hybridize to MG amplicon
A kit for detecting a nucleic acid of Mycoplasma genitalium comprising a fluorescent detectably labeled probe consisting of a third oligonucleotide sequence selected from SEQ ID NOs: 68, 71-88 and 89, wherein the detectably labeled probe is configured to hybridize to an amplicon generated by the first primer and the second primer.
Forward primer selected from defined SEQ ID NO set
A kit for detecting a nucleic acid of Mycoplasma genitalium comprising a forward primer consisting of a first oligonucleotide sequence selected from the group consisting of SEQ ID NOs: 48, 49, 53, 54, 55 and 56.
Reverse primer selected from defined SEQ ID NO set
A kit for detecting a nucleic acid of Mycoplasma genitalium comprising a reverse primer consisting of a second oligonucleotide sequence selected from the group consisting of SEQ ID NOs: 58, 62, 63, 64, 65 and 66.
Across the independent claim, the main inventive features are the defined SEQ ID NO selections for a forward primer, a reverse primer, and a fluorescently detectably labeled probe, with the probe configured to hybridize to an amplicon generated by the primers.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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