Detection methods
A method of detecting the presence of Neisseria gonorrhoeae in a sample. The method involves detecting a first target sequence taken from the NGO1642 gene and/or a second target sequence taken from the NGO1012 gene. The method may involve a step of amplifying the target sequence and may involve hybridising the target sequence to a nucleic acid probe and identifying hybridisation. The method may involve simultaneous detection of other target sequences, e.g., from other pathogens.
1 . A detection cartridge comprising:
reagents for extraction of nucleic acid from a sample;
primers that amplify genetic targets from C. trachomatis and T. vaginalis to yield amplicons; and
nucleic acid probes that hybridize to the amplicons, the nucleic acid probes including a label that signals when the nucleic acid probes are hybridized to the amplicons,
wherein the primers comprise a set of forward primers comprising 12 or more contiguous nucleotides within SEQ ID NO: 18 and 12 or more contiguous nucleotides within SEQ ID NO: 23.
2 . The cartridge of claim 1 , wherein the label is an electrochemical label.
3 . The cartridge of claim 2 , wherein the electrochemical label is a ferrocene label.
4 . The cartridge of claim 1 , wherein at least one of the primers comprises at least one modified nucleotide.
5 . The cartridge of claim 1 , wherein the cartridge comprises:
a sample inlet comprising the reagents;
an amplification section in fluid communication with the sample inlet, the amplification section comprising the primers; and
a detection section in fluid communication with the amplification section, the detection section comprising the nucleic acid probes.
6 . The cartridge of claim 1 , wherein the set of forward primers comprises 12 to 29 contiguous nucleotides within SEQ ID NO: 18.
7 . The cartridge of claim 1 , wherein the set of forward primers comprises 12 to 28 contiguous nucleotides within SEQ ID NO: 23.
8 . The cartridge of claim 1 , wherein the primers further comprise a reverse primer comprising 12 to 26 contiguous nucleotides within SEQ ID NO: 19.
9 . The cartridge of claim 1 , wherein the primers further comprise a reverse primer comprising 12 to 26 contiguous nucleotides within SEQ ID NO: 24.
10 . The cartridge of claim 1 , wherein the genetic targets include a sequence present in SEQ ID NO: 15 or SEQ ID NO: 16.
11 . The cartridge of claim 1 , wherein the genetic targets include a sequence present in SEQ ID NO: 20 or SEQ ID NO: 21.
12 . The cartridge of claim 1 , the set of primers further comprises primers that amplify a genetic target from Neisseria gonorrhoeae to yield amplicons using forward primer comprising 10 or more contiguous nucleotides within SEQ ID NO: 4 or SEQ ID NO: 9.