JPH06225800A - Oligonucleotide for detecting human alpha herpes virus, method for detecting human alpha herpes virus and reagent kit for detection - Google Patents

Abstract

PURPOSE:To simply, rapidly and specifically obtain oligonucleotides capable of detecting human alpha herpes virus in high sensitivity. CONSTITUTION:The oligonucleotides for in common detecting human alpha herpes viruses, having nucleic acid sequences shown in specific sequence tables and sequence numbers or their complementary sequences, the method for detecting human alpha herpes viruseses using these oligonucleotides as labeled probes or nucleic acid primers and the reagent kit for detection.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an oligonucleotide for conveniently and rapidly detecting human α-herpesvirus, a method for detecting human α-herpesvirus and a reagent kit for the detection.

[0002]

2. Description of the Related Art With respect to human α-herpes infection, since effective antiviral agents have been developed in recent years, the importance of early diagnosis from a small amount of patient material is increasing. Human α-herpesvirus includes herpes simplex virus type 1 (HSV-
1), the same type 2 (HSV-2), and varicella-zoster virus (VZV) are known, and the symptoms of diseases caused by these viruses may be characteristic of the virus. However, there are some symptoms in which the causative virus cannot be easily determined. In addition, these viruses are known to be latently infected after the initial infection, and cause recurrence when the immunity is lowered. Also in this case, it is difficult to identify the causative virus only from the symptoms. In the conventional diagnosis of human α-herpes, isolation of the causative virus by cell culture has been the most reliable, but with this method, it takes several days to several weeks to culture and identify. On the other hand, as a rapid diagnostic method, there is also a diagnostic method utilizing an immune reaction, but it is not sufficient in terms of sensitivity and is a characteristic of viruses belonging to the herpesviridae. Can't get In recent years, detection by a DNA diagnostic method has been attempted, but most of them are specific to each virus. To test all human α-herpesviruses at the same time, prepare respective kits and separately. It needs to be investigated and extra samples are required. A method of amplifying HSV-1 and HSV-2 with a common primer and detecting each with a specific probe has also been announced (Piiparinen and
Vaheri, Virology, 119: 27.
5, 1991), a system in which all human α-herpesviruses are commonly amplified and then measured by a probe has not been known so far.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a human α-herpes common detection oligonucleotide and a human α-herpes-specific detection oligonucleotide which can detect human α-herpesvirus simply, rapidly, specifically and with high sensitivity. Is to provide.

[0004]

[Means for Solving the Problems] As a result of various studies on the human α-herpesvirus gene, the present inventors have focused on the sequence of the DNA polymerase gene, and because of its similarity and specificity, simple, rapid, The present invention has been completed by obtaining an oligonucleotide having a sequence capable of specifically and highly sensitively detecting human α-herpesvirus, establishing a detection method using it.

That is, the present invention is an oligonucleotide for common detection of human α-herpesvirus, which has high homology to all human α-herpesviruses. Specifically, human α characterized by having the nucleic acid sequences shown in Sequence Listing or SEQ ID NOS: 1 to 4 or having complementary sequences thereof.
It is an oligonucleotide for herpes common detection.

Further, the present invention is an oligonucleotide for human α-herpesvirus-specific detection which is specific to any one of human α-herpesviruses. Specifically, human α having the nucleic acid sequences shown in Sequence Listing / SEQ ID NOS: 5 to 13 or having complementary sequences thereof
It is an oligonucleotide for detecting herpes specificity.

Furthermore, the present invention uses the oligonucleotides for common detection of human α-herpesvirus having the nucleic acid sequences shown in the sequence listing / SEQ ID NOS: 1 to 4 or having complementary sequences thereof as a nucleic acid primer or labeled. A method for detecting human α-herpesvirus in a sample, which comprises hybridizing the obtained labeled nucleic acid primer with DNA or RNA in the sample, extending the primer, and measuring the obtained primer extension product.

In the present invention, the human α-herpesvirus-specific detection oligonucleotide having the nucleic acid sequence shown in SEQ ID NO: 5 to 13 or having a complementary sequence thereof is used as a nucleic acid primer or labeled. The method for detecting human α-herpesvirus in a sample is characterized in that a labeled nucleic acid primer obtained by hybridization is hybridized with DNA or RNA in the sample, the primer is extended, and the obtained primer extension product is measured.

In the present invention, a human α-herpesvirus common detection oligonucleotide having the nucleic acid sequences shown in SEQ ID NOS: 1 to 4 or having complementary sequences thereof is subjected to amplification reaction as a nucleic acid primer, and Row table
Human α in a sample, characterized by using a human α herpesvirus-specific detection oligonucleotide having a nucleic acid sequence shown in SEQ ID NOs: 5 to 13 or having a complementary sequence thereof as a probe to identify a viral species It is a method for detecting herpes virus.

The present invention has an oligonucleotide for common detection of human α-herpesvirus having the nucleic acid sequences shown in the sequence listing / SEQ ID NOS: 1 to 4 or having complementary sequences thereof and / or the sequence listing / SEQ ID NOS: 5 to 13 A reagent kit for detecting human α-herpesvirus in a sample, which comprises an oligonucleotide for human-α-herpesvirus-specific detection, which has the nucleic acid sequence shown in or has a complementary sequence thereof.

[0011] The oligonucleotide for common detection of human α-herpesvirus of the present invention is an oligonucleotide having a high homology to all human α-herpesvirus in the gene of human α-herpesvirus, specifically, Sequence Listing / SEQ ID NO: 1 to 4 (wherein A represents adenine, C represents cytosine, G represents guanine, T represents thymine, and T at any position may be replaced with uracil (U)). Or having sequences complementary to them.

The human α-herpesvirus-specific detection oligonucleotide of the present invention is an oligonucleotide that is specific to any one of the human α-herpesviruses in the gene of human α-herpesvirus. Has the nucleic acid sequences shown in Sequence Listing or SEQ ID NOs: 5 to 13 or has a sequence complementary thereto.

The method for detecting human α-herpesvirus in a sample according to the present invention is carried out by labeling the above-mentioned oligonucleotide for detecting human α-herpesvirus, hybridizing the obtained labeled nucleic acid probe with DNA or RNA in the sample, and binding with the hybrid. This is done by measuring the body label.

Alternatively, the above-mentioned oligonucleotide for detecting human α-herpesvirus is used as a nucleic acid primer as it is, or a labeled primer obtained by labeling is hybridized with DNA or RNA in a sample, and the primer is extended to obtain the obtained primer. This is done by measuring the extension.

Hereinafter, these detection methods will be described in detail. The method of detecting human α-herpesvirus using the oligonucleotide of the present invention includes (1) a method of detecting the above-mentioned oligonucleotide as a probe, or (2) an extension reaction using the above-mentioned oligonucleotide as a primer and a DNA polymerase or the like. Then there is a way to detect. In this case, detection can be easily performed by introducing a label such as an antigen, a hapten, an enzyme, a fluorescent substance, a luminescent substance, an enzyme substrate, a radioactive substance, an insoluble carrier into the above oligonucleotide.

When the oligonucleotide of the present invention is used as a probe, it can be achieved by detecting the above-mentioned label by an appropriate detection method after hybridization with a sample.

On the other hand, when the oligonucleotide of the present invention is used as a primer, gene amplification by a DNA polymerase or the like (PCR; JP-A-62-281, Cetus) is carried out to specifically target only the human α-herpesvirus gene. Can be amplified. In PCR, human α-herpesvirus can be easily detected by incorporating a radiolabeled nucleotide during the reaction or by fractionating the amplification product by electrophoresis and detecting a specific band. Further, the amplification product can be directly detected by using the above-mentioned labeled oligonucleotide as a primer.

In any of the above methods (1) and (2), a human α-herpesvirus common oligonucleotide having a nucleic acid sequence shown in Sequence Listing or SEQ ID NOS: 1 to 4 or having a complementary sequence thereof is used. If used, the presence of any of the human alpha herpesviruses can be known.
Similarly, when a human α-herpes specific detection oligonucleotide having a nucleic acid sequence shown in Sequence Listing or SEQ ID NOs: 5 to 13 or having a complementary sequence thereof, a specific virus can be specifically detected. it can.

Further, if the amplification reaction is carried out with the common human α-herpesvirus oligonucleotide and then the specific probe is used for measurement, it is possible to detect the virus in the sample with one amplification reaction and with higher sensitivity. Become. Conversely, it is also possible to perform selection with a specific oligonucleotide and then amplify with a common oligonucleotide.

In the present invention, the method for detecting human α-herpesvirus using these oligonucleotides is different only in the means as to whether the above-mentioned oligonucleotide is used as a probe or a primer.
The detection of human α-herpesvirus is essentially the same.

[0021] In general, since oligonucleotides can be prepared by chemical synthesis, it is possible to obtain oligonucleotides of constant quality easily, in large quantities and at a low cost, as compared with cloned oligonucleotides or polynucleotides.
The oligonucleotide of the present invention is a deoxyribonucleic acid (DN
It may be A) or ribonucleic acid (RNA). In the case of ribonucleic acid, it goes without saying that the thymidine residue (T) is read as the uridine residue (U). In addition, in the synthesis, T at any position was changed to U to perform the synthesis, and D containing a uridine residue was used.
It may be NA. Similarly, it may be RNA containing a thymidine residue in which U at any position is changed to T. The oligonucleotide may have point mutations such as deletion, insertion or substitution, and modified nucleotides. Also, as mentioned above, radioactive substances, enzymes, fluorescent substances,
A known label such as a luminescent substance can be used. The labeling method may be end labeling or labeling in the middle of the sequence.
It may also be a label on the sugar, phosphate group or base moiety.

The oligonucleotide of the present invention can be bound to a solid phase carrier and used as a capture probe. In this case, a sandwich assay may be performed with a combination of a capture probe and a labeled probe, or there is a method of labeling and capturing a target nucleic acid. There is also a method in which an oligonucleotide is labeled with biotin, and after hybridization, it is captured with an avidin-binding carrier. In the sandwich assay, if the oligonucleotide of the present invention is used for either one, specific measurement can be performed with the present oligonucleotide, and there is no problem even if the specificity of the other oligonucleotide is slightly low.

The reagent kit for detecting human α-herpesvirus of the present invention contains the above-mentioned oligonucleotide for common detection of human α-herpesvirus and / or the oligonucleotide for specific detection of human α-herpesvirus. Further, if necessary, a DNA polymerase, dNTP, an enzyme buffer and a substance for detecting a label are included.

[0024]

EXAMPLES The present invention will be specifically described below with reference to examples. Example 1 (Synthesis of Various Oligonucleotides) Using ABI DNA Synthesizer Model 391, various oligonucleotides having the sequences shown in the Sequence Listing and SEQ ID NOs: 1 to 13 were synthesized by the phosphoamidite method. Hereinafter, the various oligonucleotides shown in Sequence Listing and SEQ ID NOS: 1 to 13 are referred to as oligonucleotides (1) to (13), respectively. Method is ABI
According to the company manual, it was carried out on a 0.2 mM scale.
The deprotection of various oligonucleotides is 55 with ammonia water.
C was carried out overnight. Purification was carried out by FPLC manufactured by Pharmacia on a reverse phase column. If necessary, the synthesized oligonucleotide had a ³² P-phosphate group bonded to the 5'end by the following method.

Reaction solution composition Oligonucleotide 5 to 20 picomoles 10 × T4 polynucleotide kinase buffer 10 μl 1 mM [γ-32P ATP (10 mCi / ml) 1 μl T4 polynucleotide kinase (manufactured by Toyobo) 10 units

The reaction mixture prepared as described above was placed at 37 ° C.
And reacted for 1 hour. Here, 10 × T4 polynucleotide kinase buffer is 0.5M Tris-HCl (pH8.0), 0.1MM
gCl ₂ , 0.1M 2-mercaptoethanol.

Example 2 (Common Amplification of Human α-Herpesvirus) (1) Kit A: Kit for amplifying human α-herpesvirus-derived nucleic acid (a) Oligonucleotide of Example 1 (1) (b) Example Oligonucleotide 1 (2) (c) Oligonucleotide of Example 1 (3) (d) Oligonucleotide of Example 1 (4) (e) Tth DNA polymerase (manufactured by Toyobo) dATP, dCTP, dGTP, dTTP

(2) Preparation of viral nucleic acid Herpes simplex virus type 1 (HSV-1; Human)
For culturing siebert strain as a standard strain of simplex virus type 1), UW268 strain as a standard strain of herpes simplex virus type 2 (HSV-2), and kawaguchi strain as a standard strain of varicella-zoster virus (VZV). I was there. In addition, in order to confirm the specificity, EB virus (EBV; Epstein-Barr virus)
s) B95-8 strain, human cytomegalovirus (CM
V; cytomegalovirus) AD169 strain,
Human herpesvirus type 6 (HHV-6) type A (U11
02) strain, type B (Hashimoto strain), and human placenta DNA
(Manufactured by Sigma) was used.

5 μl of the nucleic acid solution described above and 1 μl each of the oligonucleotides (1) to (3) of Example 1 or (2) and (4) of Example 1 were added to 93 μl of the PCR reaction solution as a primer to prepare a nucleic acid derived from human α-herpesvirus. Amplification was performed. The composition of the reaction solution is as follows.

Composition of reaction solution KCl 50 mM Tris-HCl (pH 8.3) 10 mM MgCl ₂ 1.5 mM gelatin 0.01% dATP, dCTP, dGTP, dTTP 0.2 mM Tth DNA polymerase 40 units / ml

The reaction conditions are as follows. Thermal denaturation: 94 ° C., 1 minute, annealing: 50 ° C., 2 minutes Polymerization reaction: 72 ° C., 1.5 minutes 30 times. These operations are Perkin-Elmer /
It was performed using a DNA thermal cycler manufactured by Cetus (Perkin-Elmer / Cetus).

Detection 10 μl of the reaction solution was electrophoresed on a 2% agarose gel and stained with ethidium bromide, and then fluorescence from ultraviolet rays was detected (FIG. 1). In the figure, (A) is the oligonucleotide (1)
And (2) are used as primers, and (B) shows the case where oligonucleotides (3) and (4) are used. Lane 2
9 is HSV-1, HSV-2, VZV, EB, respectively.
V, CMV, HHV-6 (A), HHV-6 (B), and human placental DNA are shown. Lanes 1 and 10 show molecular weight markers. The electrical conditions for electrophoresis are constant voltage 10
The time was 0 V and the time was 30 minutes. Operating method and other conditions
The method described in Maniatis et al., Molecular Cloning (1982) was followed. In addition to the reaction solution, molecular weight markers are also electrophoresed at the same time,
The length of the detected nucleotide fragment was calculated by comparing the relative mobilities.

Results In the PCR product, a band was observed only when human α-herpesvirus was used (lanes 1-3). From the molecular weight marker, its size is 260 base pairs (A) when oligonucleotides (1) and (3) are used.
When using (2) and (4), it was identified as 330 base pairs (B). This was in good agreement with the nucleotide length calculated from the nucleic acid sequence.

Example 3 (Specific detection of human α-herpesvirus) In order to identify which human α-herpesvirus the amplification product of Example 2 was derived from, dot blot hybridization was carried out.

(1) Kit B: Kit for detecting human α-herpesvirus-derived nucleic acid Oligonucleotides (5) to (13) of Example 1 having a ³² P-phosphate group at the 5'end ing. (2) Dot Blot Hybridization 100 μl of 0.3 N NaOH was added to 1 μl of the PCR product of Example 2, denatured at room temperature for 15 minutes, and wetted with 5 × SSC using a dot blotter (manufactured by BRL). It was blotted on a nylon membrane (High Bond N Plus, manufactured by Amersham). SSC is a 15 mM trisodium citrate and 15 mM sodium chloride solution, and 5 × SSC is a 5-fold concentrated solution of SSC. This membrane was heated at 80 ° C. for 30 minutes to immobilize the nucleic acid, and then hybridized. First, the dried membrane (8 × 12 cm) was immersed in 5 × SSC for 5 minutes. Next, transfer the membrane to a hybridization bag (manufactured by BRL), add 5 ml of hybridization buffer (5 × SSC, 0.5% bovine serum albumin, 0.5% polyvinylpyrrolidone, 1% sodium dodecyl sulfate) to the policy. Seal with a stirrer and pre-hybridize at 50 ° C. for 15 minutes. Next, the oligonucleotide probe of kit B
Hybridization was carried out at 55 ° C. for 15 minutes with 5 ml of a hybridization buffer containing 5 μl of any of (5) to (13). The membrane was taken out from Polyvac and washed with Washing Solution-1 (1 × SSC, 1% sodium dodecyl sulfate) at 55 ° C. for 5 minutes with shaking twice. The membrane was thoroughly dried after it was washed with Shake Solution-2 (1 × SSC) twice at room temperature for 5 minutes with shaking.

(3) Detection An X-ray film (New AIF RX manufactured by Fuji Photo Industry Co., Ltd.) was adhered to the nylon film and exposed at -80 ° C for a whole day and night. Human α-herpesvirus was detected from the photosensitivity of the film.

Results The oligonucleotide probe specific to each virus specifically reacts only with the amplification product derived from each virus, and no reaction with the amplification products derived from other viruses or human DNA was observed. It was

Example 4 Detection of Human α-Herpes Virus Using Patient Specimens It was examined whether the oligonucleotides of the present invention are effective for detecting virus derived from patients. At the same time, the results were compared using a commercially available immunoassay kit (manufactured by Syva).

(1) Kit A: Kit for amplifying human α-herpesvirus-derived nucleic acid (a) Oligonucleotide of Example 1 (1) (b) Oligonucleotide of Example 1 (2) (c) Example Oligonucleotide 1 (3) (d) Oligonucleotide 1 of Example 1 (4) (e) Tth DNA polymerase (manufactured by Toyobo) dATP, dCTP, dGTP, dTTP Kit B: kit for detecting human α-herpesvirus-derived nucleic acid The oligonucleotides (5) to (13) of Example 1 have a ³² P-phosphate group at the 5'end.

(2) Preparation of Specimen From a virus culture solution isolated from a patient diagnosed with herpes simplex virus infection, the method of Kido et al.
nic. Microbiol. 29; 76,1991).
Viral nucleic acid was extracted according to.

(3) Amplification and Detection Using the amplification oligonucleotides (1) and (4), and the detection oligonucleotides (5), (8) and (13), Example 2 and Example 3 were used. Amplification and detection of the virus in the sample were performed in the same manner as in.

(4) Detection by immunoassay The virus in the sample was detected according to the instructions attached to the kit of Syva. Results Of the 16 patient isolates examined, 14 were HSV1 and
There were two HSV2 strains. This was in good agreement with the result of detection by a commercial immunoassay (Table 1).

[0043]

[Table 1]

[0044]

INDUSTRIAL APPLICABILITY According to the present invention, it has become possible to measure human α-herpesvirus, which has conventionally been time-consuming to culture, quickly, simply and specifically with high sensitivity. The oligonucleotide of the present invention also serves as a primer for an amplification reaction,
It can also be used as a probe for direct detection. In particular, by effectively using the common detection oligonucleotide and the specific detection oligonucleotide,
All human α-herpesviruses can be detected with high sensitivity even from batch samples, and their clinical significance is great.

[0045]

[Sequence list]

SEQ ID NO: 1 Sequence length: 20 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..20 Method of determining features : S Other features: Sequence common to the DNA polymerase gene of human α-herpesvirus. Array CCAAGTATCA TTCAGGCCCA 20

SEQ ID NO: 2 Sequence length: 20 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..20 Method determined: S Other features: Sequence common to the DNA polymerase gene of human α-herpesvirus. Sequence GTGGTGGTAT TTGATTTTGC 20

SEQ ID NO: 3 Sequence length: 20 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..20 Method determined: S Other features: Sequence common to the DNA polymerase gene of human α-herpesvirus. Sequence ATCGCGGCCT GTTGTTTGTC 20

SEQ ID NO: 4 Sequence length: 20 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..20 Method determined: S Other features: Sequence common to the DNA polymerase gene of human α-herpesvirus. Array ACTCCTGTGA AACCGTACAC 20

SEQ ID NO: 5 Sequence length: 16 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..16 Features Method determined: S Other characteristics: having a sequence specific to the sequence of the DNA polymerase gene of HSV-1. Sequence AGGCGGGCAA GGACTA 16

SEQ ID NO: 6 Sequence length: 16 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..16 Features Method determined: S Other characteristics: having a sequence specific to the sequence of the DNA polymerase gene of HSV-1. Sequence TCGCACGTGA GCCTTG 16

SEQ ID NO: 7 Sequence length: 15 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..15 Method determined: S Other characteristics: having a sequence specific to the sequence of the DNA polymerase gene of HSV-1. Sequence AGGGCCGACG CAGTG 15

SEQ ID NO: 8 Sequence length: 16 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..16 Method determined: S Other characteristics: having a sequence specific to the sequence of the DNA polymerase gene of HSV-2. Sequence AGGCGGACCG GGACTA 16

SEQ ID NO: 9 Sequence length: 16 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..16 Method determined: S Other characteristics: having a sequence specific to the sequence of the DNA polymerase gene of HSV-2. Sequence GCGTACGTGG GCCTTC 16

SEQ ID NO: 10 Sequence length: 15 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..22 Method determined: S Other characteristics: having a sequence specific to the sequence of the DNA polymerase gene of HSV-2. Sequence AGAGCCTGCT GAGCATCCTG CT 22

SEQ ID NO: 11 Sequence length: 27 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..27 Method determined: S Other features: having a sequence specific for the sequence of the VZV DNA polymerase gene. Sequence GACGGTTAAA CGTTTGAATC CATCCGA 27

SEQ ID NO: 12 Sequence length: 26 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..26 Features Method determined: S Other features: having a sequence specific for the sequence of the VZV DNA polymerase gene. Sequence TTTTACCACG CTAACGTTAA ATTTTG 26

SEQ ID NO: 13 Sequence length: 23 Sequence type: Nucleic acid Number of strands: Both forms Topology: Linear Sequence type: Other nucleic acid Synthetic DNA Sequence features Location: 1..23 Features Method determined: S Other features: having a sequence specific for the sequence of the VZV DNA polymerase gene. Sequence TTATGCCACC TTTACAGTTG GAG 23

[Brief description of drawings]

FIG. 1 shows the results of electrophoresis of Example 2. In the figure, (A)
Shows the case where oligonucleotides (1) and (2) were used as primers, and (B) shows the case where oligonucleotides (3) and (4) were used. Lanes 2-9 are HSV-1 and HS, respectively.
V-2, VZV, EBV, CMV, HHV-6 (A),
HHV-6 (B) and human placental DNA are shown.

FIG. 2 shows an X-ray film of Example 3. In the figure, A,
B, C, D, E, F, G, H, and I are oligonucleotides (5), (6), (7), (8), (9), (10), (11), and (1), respectively.
The results of using 2) and (13) as probes are shown. Again 2
9 to 9 correspond to 2 to 9 in the second embodiment.

Claims (8)

[Claims] 1. An oligonucleotide for common detection of human α-herpesvirus, which has a high homology to all human α-herpesviruses. 2. The human α-herpes common detection oligonucleotide according to claim 1, which has the nucleic acid sequences shown in Sequence Listing or SEQ ID NOS: 1 to 4 or has a complementary sequence thereof. 3. An oligonucleotide for human α-herpesvirus-specific detection which is specific for any one of human α-herpesviruses. 4. The human α-herpes specific detection oligonucleotide according to claim 3, which has the nucleic acid sequences shown in SEQ ID NOS: 5 to 13 or has a complementary sequence thereof. 5. The human α-herpesvirus common detection oligonucleotide according to claim 1 is used as a nucleic acid primer as it is, or a labeled nucleic acid primer obtained by labeling is hybridized with DNA or RNA in a sample to extend the primer. And a method for detecting human α-herpesvirus in a sample, which comprises measuring the obtained primer extension product. 6. The human α-herpesvirus-specific detection oligonucleotide according to claim 3 is used as a nucleic acid primer as it is, or a labeled nucleic acid primer obtained by labeling is hybridized with DNA or RNA in a sample to extend the primer. And a method for detecting human α-herpesvirus in a sample, which comprises measuring the obtained primer extension product. 7. The human α-herpesvirus common detection oligonucleotide according to claim 1 is used as a nucleic acid primer for amplification reaction, and then the human α-herpesvirus-specific detection oligonucleotide according to claim 3 is used as a probe. A method for detecting human α-herpesvirus in a sample, which comprises: 8. An oligonucleotide for common detection of human α-herpesvirus according to claim 1, and / or claim 3.
A reagent kit for detecting human α-herpesvirus in a sample, which comprises the oligonucleotide for detecting specific human α-herpesvirus.