Neisseria gonorrhoeae detection

Abstract

A method for determining whether a human individual is or has been infected with Neisseria gonorrhoeae, is provided. The method detects a Neisseria gonorrhoeae, porA nucleic acid fragment obtained from a biological sample. The method includes subjecting the biological sample to nucleic acid sequence amplification using primers having respective nucleotide sequences according to SEQ ID NO:1 and SEQ ID NO:2, to thereby produce a porA Neisseria gonorrhoeae, amplification product. The amplification product is detected by fluorescence resonance energy transfer using oligonucleotides having respective nucleotide sequences according to SEQ ID NO:3 which has a donor fluorophore and SEQ ID NO:4, which has an acceptor fluorophore.

1 . A method of determining whether an individual is infected with  Neisseria gonorrhoeae , said method including the step of detecting an isolated porA nucleic acid of  Neisseria gonorrhoeae , if present in a biological sample obtained from said individual, a presence of said porA nucleic acid indicating that said individual is infected with  Neisseria gonorrhoeae , wherein said isolated porA nucleic acid comprises a nucleotide sequence of an amplification product obtainable by nucleic acid sequence amplification using primers having a nucleotide sequence selected from the group consisting of: SEQ ID NO:1 and SEQ ID NO:2. 
     
     
         2 . The method of  claim 1 , wherein the nucleotide sequence of the amplification product comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:3; SEQ ID NO:4; a nucleotide sequence complementary to SEQ ID NO:3; and a nucleotide sequence complementary to SEQ ID NO:4. 
     
     
         3 . The method of  claim 1 , wherein the nucleotide sequence is at least 80% identical to residues 681-812 of SEQ ID NO:10. 
     
     
         4 . The method of  claim 1 , wherein said method includes the step of distinguishing said isolated porA nucleic acid of  Neisseria gonorrhoeae  from a porA nucleic of  Neisseria meningitidis  present in said biological sample. 
     
     
         5 . The method of  claim 1 , further including the step of using one or more oligonucleotide probes for detecting said amplification product by probe hybridization, wherein the probe comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:3 and SEQ ID NO:4. 
     
     
         6 . The method of  claim 5 , wherein detection of said amplification product is performed using fluorescence resonance energy transfer (FRET). 
     
     
         7 . The method of  claim 1 , including the step of subjecting the amplification product to nucleotide sequencing. 
     
     
         8 . A method of determining whether an individual is infected with  Neisseria gonorrhoeae , said method including the step of detecting an isolated porA nucleic acid of  Neisseria gonorrhoeae , if present in a biological sample obtained from said individual, a presence of said porA nucleic acid indicating that said individual is infected with  Neisseria gonorrhoeae , wherein said isolated porA nucleic acid comprises a nucleotide sequence of an amplification product produced by nucleotide sequence amplification, wherein the nucleotide sequence is at least 80% identical to residues 681-812 of SEQ ID NO:10. 
     
     
         9 . The method of  claim 8 , wherein the amplification product is produced by nucleotide sequence amplification using primers having a nucleotide sequence selected from the group consisting of: SEQ ID NO:1 and SEQ ID NO:2. 
     
     
         10 . The method of  claim 8 , further including the step of using one or more oligonucleotide probes for detecting said amplification product by probe hybridization, wherein the probes comprise respectively a nucleotide sequence selected from the group consisting of SEQ ID NO:3 and SEQ ID NO:4. 
     
     
         11 . The method of  claim 10 , wherein detection of said amplification product is performed using fluorescence resonance energy transfer (FRET). 
     
     
         12 . The method of  claim 8 , including the step of subjecting the amplification product to nucleotide sequencing. 
     
     
         13 . An oligonucleotide comprising a nucleotide sequence according to any one of SEQ ID NOS:1-9, or a nucleotide sequence complementary thereto. 
     
     
         14 . The oligonucleotide of  claim 8 , further comprising a label. 
     
     
         15 . A kit for detecting a porA nucleic acid of  Neisseria gonorrhoeae  comprising one or more oligonucleotides according to  claim 13  and a DNA polymerase and/or one or more detection reagents.