Detection of herpes simplex virus

A method of distinguishing between HSV-1 and HSV-2 in a biological sample from an individual, the method comprising:

• performing at least one cycling step, wherein said cycling step comprises an amplifying step and a hybridizing step, wherein said amplifying step comprises contacting said sample with a pair of HSV DNA polymerase primers to produce an HSV-1 DNA polymerase amplification product if a nucleic acid molecule encoding HSV-1 DNA polymerase is present in said sample and/or an HSV-2 DNA polymerase amplification product if a nucleic acid molecule encoding HSV-2 DNA polymerase is present in said sample, wherein said HSV-1 and said HSV-2 amplification products differ in sequence by at least one nucleotide, wherein said hybridizing step comprises contacting said sample with a pair of HSV DNA polymerase probes, wherein the members of said pair of HSV DNA polymerase probes hybridize within no more than five nucleotides of each other, wherein a first HSV DNA polymerase probe of said pair of HSV DNA polymerase probes is labeled with a donor fluorescent moiety and wherein a second HSV DNA polymerase probe of said pair of HSV DNA polymerase probes is labeled with a corresponding acceptor fluorescent moiety;
• detecting the presence or absence of FRET between said donor fluorescent moiety of said first HSV DNA polymerase probe and said corresponding acceptor fluorescent moiety of said second HSV DNA polymerase probe, wherein the presence of FRET is indicative of the presence of HSV in said biological sample, wherein the absence of FRET is indicative of the absence of HSV in said biological sample;
• and
• determining the melting temperature between one or both of said HSV DNA polymerase probes and said HSV DNA polymerase amplification products, wherein said melting temperature distinguishes between HSV-1 and HSV-2.

A method for detecting the presence or absence of HSV in a biological sample from an individual, said method comprising:

• performing at least one cycling step, wherein said cycling step comprises an amplifying step and a hybridizing step, wherein said amplifying step comprises contacting said sample with a pair of HSV DNA polymerase primers to produce an HSV DNA polymerase amplification product if a nucleic acid molecule encoding HSV DNA polymerase is present in said sample, wherein said hybridizing step comprises contacting said sample with an HSV DNA polymerase probe, wherein said HSV DNA polymerase probe is labeled with a donor fluorescent moiety and a corresponding acceptor fluorescent moiety;
• and
• detecting the presence or absence of fluorescence resonance energy transfer (FRET) between said donor fluorescent moiety and said corresponding acceptor fluorescent moiety of said HSV DNA polymerase probe,
• wherein the presence or absence of FRET is indicative of the presence or absence of HSV in said sample.

A method for detecting the presence or absence of HSV in a biological sample from an individual, said method comprising:

• performing at least one cycling step, wherein said cycling step comprises an amplifying step and a dye-binding step, wherein said amplifying step comprises contacting said sample with a pair of HSV DNA polymerase primers to produce an HSV DNA polymerase amplification product if a nucleic acid molecule encoding HSV DNA polymerase is present in said sample, wherein said dye-binding step comprises contacting said HSV DNA polymerase amplification product with a nucleic acid binding dye;
• and
• detecting the presence or absence of binding of said nucleic acid binding dye to said amplification product,
• wherein the presence of binding is indicative of the presence of HSV in said sample, and wherein the absence of binding is indicative of the absence of HSV in said sample.