Rapid and sensitive detection of herpes simplex virus

A method for detecting the presence or absence of herpes simplex virus, wherein the method comprises the steps of:

• (a) lysing the cells in a specimen to release nucleic acid molecules;
• (b) contacting the nucleic acid molecules with oligonucleotide primer pair SEQ ID NO:7 and SEQ ID NO:8 under suitable conditions permitting hybridization of the oligonucleotides to the nucleic acid molecules;
• (c) enzymatically amplifying a specific region of the nucleic acid molecules comprising target sequences of the herpes simplex virus polymerase gene using oligonucleotides SEQ ID NO:7 and SEQ ID NO:8 as primers;
• (d) contacting amplified target sequences from step (c), if present, with an oligonucleotide probe selected from the group consisting of SEQ ID NO:11, or SEQ ID NO:12, labeled with a detectable moiety under suitable conditions permitting liquid hybridization of the labeled oligonucleotide probe to amplified target sequences;
• and
• (e) detecting the presence of amplified target sequences by detecting the detectable moiety of the labeled oligonucleotide probe hybridized to amplified target sequences in a gel retardation assay;
• wherein the presence of the amplified target sequences hybridized to labeled oligonucleotide probe SEQ ID NO:11 correlates to the presence of herpes simplex virus in the specimen;
• and wherein the presence of the amplified target sequences hybridized to labeled oligonucleotide probe SEQ ID NO:12 correlates to the presence of herpes simplex virus 2 in the specimen with a sensitivity of detection at least as low as 10 TCID.sub.50.

A method for detecting the presence or absence of herpes simplex virus, wherein the method comprises the steps of:

• (a) lysing the cells in a specimen to release nucleic acid molecules;
• (b) contacting the nucleic acid molecules with oligonucleotide primer pair SEQ ID NO:9 and SEQ ID NO:10 under suitable conditions permitting hybridization of the oligonucleotides to the nucleic acid molecules;
• (c) enzymatically amplifying a specific region of the nucleic acid molecules comprising target sequences of the herpes simplex virus thymidine kinase gene using oligonucleotides SEQ ID NO:9 and SEQ ID NO:10 as primers;
• (d) contacting amplified target sequences from step (c), if present, with an oligonucleotide probe selected from the group consisting of SEQ ID NO:13, or SEQ ID NO:14, labeled with a detectable moiety under suitable conditions permitting liquid hybridization of the labeled oligonucleotide probe to amplified target sequences;
• and
• (e) detecting the presence of amplified target sequences by detecting the detectable moiety of the labeled oligonucleotide probe hybridized to amplified target sequences in a gel retardation assay;
• wherein the presence of the amplified target sequences hybridized to labeled oligonucleotide probe SEQ ID NO:13 correlates to the presence of herpes simplex virus 1 in the specimen;
• and wherein the presence of the amplified target sequences hybridized to labeled oligonucleotide probe SEQ ID NO:14 correlates to the presence of herpes simplex virus in the specimen with a sensitivity of detection at least as low as 10 TCID.sub.50.

A diagnostic test kit for detection of HSV comprising, in separate packaging:

• (a) at least one oligonucleotide primer pair selected from the group consisting of SEQ ID NO:7 & SEQ ID NO:8, SEQ ID NO:9 & SEQ ID NO:10, and both SEQ ID NO:7 & SEQ ID NO:8 and SEQ ID NO:9 & SEQ ID NO:10;
• and
• (b) at least one oligonucleotide probe labeled with a detectable moiety selected from the group consisting of SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, and SEQ ID NO:14;
• wherein SEQ ID NO:11 and SEQ ID NO:12 is for use with primer pair SEQ ID NO:7 & SEQ ID NO:8, and wherein SEQ ID NO:13 and SEQ ID NO:14 is for use with primer pair SEQ ID NO:9 & SEQ ID NO:10.

A method for detecting the presence or absence of herpes simplex virus, wherein the method comprises the steps of:

• (a) lysing the cells in a specimen to release nucleic acid molecules;
• (b) contacting the nucleic acid molecules with oligonucleotide primer pair SEQ ID NO:7 and SEQ ID NO:8 under suitable conditions permitting hybridization of the oligonucleotides to the nucleic acid molecules, wherein at least one of oligonucleotides selected from the oligonucleotide primer pair is labeled with a detectable moiety;
• (c) enzymatically amplifying a specific region of the nucleic acid molecules comprising target sequences of the herpes simplex virus polymerase gene using oligonucleotides SEQ ID NO:7 and SEQ ID NO:8 as primers;
• and
• (d) electrophoresing any amplified molecules present from step (c) and detecting the presence of amplified target sequences by detecting the detectable moiety in the amplified target sequences, wherein the presence of the amplified target sequences correlates to the presence of herpes simplex virus in the specimen with a sensitivity of detection at least as low as 1 TCID.sub.50.

A method for detecting the presence or absence of herpes simplex virus, wherein the method comprises the steps of:

• (a) lysing the cells in a specimen to release nucleic acid molecules;
• (b) contacting the nucleic acid molecules with oligonucleotide primer pair SEQ ID NO:9 and SEQ ID NO:10 under suitable conditions permitting hybridization of the oligonucleotides to the nucleic acid molecules, wherein at least one of oligonucleotides selected from the oligonucleotide primer pair is labeled with a detectable moiety;
• (c) enzymatically amplifying a specific region of the nucleic acid molecules comprising target sequences of the herpes simplex virus polymerase gene using oligonucleotides SEQ ID NO:9 and SEQ ID NO:10 as primers;
• and
• (d) electrophoresing any amplified molecules present from step (c) and detecting the presence of amplified target sequences by detecting the detectable moiety in the amplified target sequences, wherein the presence of the amplified target sequences correlates to the presence of herpes simplex virus in the specimen with a sensitivity of detection at least as low as 1 TCID.sub.50.