|
Rapid and sensitive detection of herpes simplex virus
| Details |
Inventors: Greenberg, Steven J.; Evans, Mary Jo;
Assignee: Health Research Inc. (Buffalo, NY)
Primary Examiner: Myers; Carla J.
Assistant Examiner:
Attorney, Agent or Firm: Hodgson, Russ, Andrews, Woods & Goodyear
The present invention relates to novel compositions comprising herpesvirus-specific oligonucleotides which are useful as primers to amplify particular regions of the genome either herpes simplex virus or cytomegalovirus during enzymatic nucleic acid amplification. The invention also provides a rapid, sensitive and specific method for the detection of the respective herpesvirus which may be present in a clinical specimen, using the herpesvirus-specific primers and enzymatic nucleic acid amplification; hybridization of amplified target sequences, if present, with one or more herpesvirus-specific oligonucleotide probes which are labeled with a detectable moiety; and detection of the detectable moiety of labeled oligonucleotide probe hybridized to amplified target sequences of herpesvirus DNA. |
|
DETAILED DESCRIPTION OF THE INVENTION 5.
1 Definitions By the terms "enzymatically amplify" or "amplify" is meant, for the purposes of the specification or claims, a process by which nucleic acid sequences are amplified in number.
There are several means for enzymatically amplifying nucleic acid sequences.
Currently the most commonly used method, PCR (polymerase chain reaction, Cetus Corporation) involves the use of a thermostable DNA polymerase, known sequences as primers, and heating cycles which separate the replicating deoxyribonucleic acid (DNA) strands and exponentially amplify a gene of interest.
Other amplification methods include LCR (ligase chain reaction, BioTechnica International) which utilizes DNA ligase, and a probe consisting of two halves of a DNA segment that is complementary to the sequence of the DNA to be amplified; enzyme QB replicase (Gene-Trak Systems) and a ribonucleic acid (RNA) sequence template attached to a probe complementary to the DNA to be copied which is used to make a DNA template for exponential production of complementary RNA; and NASBA (nucleic acid sequence-based amplification, Cangene Corporation) which can be performed on RNA or DNA as the nucleic acid sequence to be amplified.
By the term "composition" is meant, for the purposes of the specification or claims, a combination of elements which may include one or more of the following: the reaction buffer for the respective method of enzymatic amplification, plus one or more oligonucleotides specific for herpesvirus CMV or HSV-1 or HSV-2; and one or more oligonucleotides specific for herpesvirus CMV or HSV-1 or HSV-2, wherein said oligonucleotide is labeled with a detectable moiety.
By the term "complementarity" or "complementary" is meant, for the purposes of the specification or claims, a sufficient number in the oligonucleotide of complementary base pairs in its sequence to interact specifically (hybridize) with the target nucleic acid sequence of the herpesvirus to be amplified or detected.
As known to those skilled in the art, a very high degree of complementarity is needed for specificity and sensitivity involving hybridization, although it need not be 100%
|
|
Semiconductor manufacture 
