Methods for detecting and typing herpes simplex virus

Abstract

Detailed herein are methods of detecting herpes simplex virus and differentiating between types 1 and 2 by simultaneous detection of type-specific gene sequences. In particular aspects, individuals infected with HSV-1 can be distinguished from those infected with HSV-2 by amplification and detection of the HSV-1 glycoprotein B gene or the HSV-2 UL-8 gene. Primers and probes for the differential detection of HSV-1 and HSV-2 are provided.

Description

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Exemplary sequence (SEQ ID NO: 1) of a region of HSV-1 glycoprotein B cDNA (corresponding to bases 1-243 of GenBank ID g6165611) showing the preferred locations for hybridizing PCR primers (shaded regions), and a preferred location for a hybridizing probe (bold underlined).

FIG. 2. Exemplary sequence (SEQ ID NO:2) of a region of the HSV-2-UL-8 gene (numbering refers to numbering of HSV-2 genome) showing the preferred locations for hybridizing PCR primers (shaded regions), and a preferred location for a hybridizing probe (bold underlined).

Claims

1. A method for detecting and differentiating between HSV-1 and HSV-2 in a sample, said method comprising, detecting a first target sequence specific for HSV-1 when HSV-1 nucleic acids are present in said sample, and detecting a second target sequence specific for HSV-2 when HSV-2 nucleic acids are present in said sample, wherein said first target sequence and said second target sequence are from different genes, and wherein detection of said first target sequence is indicative of an HSV-1 infection and detection of said second target sequence is indicative of an HSV-2 infection.

2. A method according to claim 1, wherein said first target sequence is from the HSV-1 glycoprotein B gene.

3. A method according to claim 1, wherein said second target sequence is from the HSV-2 UL-8 gene.

4. A method according to claim 1, wherein said first target sequence is from the HSV-1 glycoprotein B gene and said second target sequence is from the HSV-2 UL-8 gene.

5. A method according to claim 1, wherein said detecting further comprises, amplifying said first and second target sequences.

6. A method according to claim 5, wherein said amplifying is accomplished with the polymerase chain reaction.

7. A method according to claim 1, wherein said detecting of said first and second target sequences occurs in the same reaction vessel.

8. A method according to claim 1, wherein said detecting of said first and second target sequences occurs in separate reaction vessels.

9. A method according to claim 1, wherein said detecting further comprises, amplifying a region of the HSV-1 glycoprotein B (gB) gene with a first pair of type-specific primers when HSV-1 nucleic acids are present in said sample, amplifying a region of the HSV-2 UL-8 gene with a second pair of type-specific primers when HSV-2 nucleic acids are present in said sample, and detecting said amplified regions with a first gene-specific probe and a second gene-specific probe.

10. A method according to claim 9, wherein said probes are labeled.

11. A method according to claim 10, wherein said probes are distinguishably labeled.

12. A method according to claim 9, wherein said first pair of type-specific primers comprises a forward primer comprising the sequence set forth in SEQ ID NO:3 and a reverse primer.

13. A method according to claim 9, wherein said first pair of type-specific primers comprises a forward primer and a reverse primer comprising the sequence set forth in SEQ ID NO:4.

14. A method according to claim 9, wherein said second pair of type-specific primers comprises a forward primer comprising the sequence set forth in SEQ ID NO:6 and a reverse primer.

15. A method according to claim 9, wherein said second pair of type-specific primers comprises a forward primer and a reverse primer comprising the sequence set forth in SEQ ID NO:7.

16. A method according to claim 9, wherein said first gene-specific probe comprises the sequence set forth in SEQ ID NO:5.

17. A method according to claim 9, wherein said second gene-specific probe comprises the sequence set forth in SEQ ID NO:8.

18. A method of diagnosing an infection in individual as being HSV-1 or HSV-2, said method comprising, detecting a first target sequence specific for HSV-1 when HSV-1 nucleic acids are present in a biological sample from said individual, and detecting a second target sequence specific for HSV-2 when HSV-2 nucleic acids are present in said sample, wherein said first target sequence and said second target sequence are from different genes, and wherein detection of said first target sequence is indicative of an HSV-1 infection and detection of said second target sequence is indicative of an HSV-2 infection.

19. A method according to claim 18, wherein said first target sequence is from the HSV-1 glycoprotein B gene.

20. A method according to claim 18, wherein said second target sequence is from the HSV-2 UL-8 gene.

21. A method according to claim 18, wherein said first target sequence is from the HSV-1 glycoprotein B gene and said second target sequence is from the HSV-2 UL-8 gene.

22. A method according to claim 18, wherein said detecting further comprises, amplifying said first and second target sequences.

23. A method according to claim 22, wherein said amplifying is accomplished with the polymerase chain reaction.

24. A method according to claim 18, wherein said detecting of said first and second target sequences occurs in the same reaction vessel.

25. A method according to claim 18, wherein said detecting of said first and second target sequences occurs in separate reaction vessels.

26. A method according to claim 18, wherein said detecting further comprises, amplifying a region of the HSV-1 glycoprotein B (gB) gene with a first pair of type-specific primers when HSV-1 nucleic acids are present in said sample, amplifying a region of the HSV-2 UL-8 gene with a second pair of type-specific primers when HSV-2 nucleic acids are present in said sample, anddetecting said amplified regions with a first gene-specific probe and a second gene-specific probe.

27. A method according to claim 26, wherein said probes are labeled.

28. A method according to claim 27, wherein said probes are distinguishably labeled.

29. A method according to claim 26, wherein said first pair of type-specific primers comprises a forward primer comprising the sequence set forth in SEQ ID NO:3 and a reverse primer.

30. A method according to claim 26, wherein said first pair of type-specific primers comprises a forward primer and a reverse primer comprising the sequence set forth in SEQ ID NO:4.

31. A method according to claim 26, wherein said second pair of type-specific primers comprises a forward primer comprising the sequence set forth in SEQ ID NO:6 and a reverse primer.

32. A method according to claim 26, wherein said second pair of type-specific primers comprises a forward primer and a reverse primer comprising the sequence set forth in SEQ ID NO:7.

33. A method according to claim 26, wherein said first gene-specific probe comprises the sequence set forth in SEQ ID NO:5.

34. A method according to claim 26, wherein said second gene-specific probe comprises the sequence set forth in SEQ ID NO:8.