Abstract: It is an object of the present invention to provide a composition for catalyzing the cleavage of a single-stranded DNA and the binding of such single-stranded DNA. The present invention provides a composition for cleaving a single-stranded DNA and/or binding the 5?-terminus of such single-stranded DNA to the 3?-terminus thereof, which comprises an Ev1 protein. Moreover, the present invention also provides a composition for cleaving a single-stranded DNA and/or binding the 5?-terminus of such single-stranded DNA to the 3?-terminus thereof, which further comprises a Rad51B protein and/or a DNA topoisomerase type I protein, as well as the Ev1 protein.

Abstract: The described method provides, methods, and kits to produce, identify, catalog and classify a comprehensive collection of nucleic acid targets produced from a nucleic acid sample. The method, referred to as Cataloging and Classification of Sequence Tags, involves generating a set of target nucleic acid fragments; coupling the target nucleic acid fragments to a nucleic acid bridge comprising, for example, two or more primer binding sites and two recognition sites for cleavage at a site offset from the recognition site to the fragment's end; and cleaving the fragments to generate chimeric nucleic acids of known length. The nucleic acid bridge is thus disposed between the two nucleic acid fragments in the chimeric nucleic acid. The resulting duplex nucleic acids comprise a set of sequence tags (i.e., by amplification using universal primers), comprising an addressable portion, a target nucleic portion and a portion of the nucleic acid bridge.

Abstract: The present invention provides a method of covalently joining a DNA strand to an RNA strand comprising (a) forming a topoisomerase-DNA intermediate by incubating a DNA cleavage substrate comprising a topoisomerase cleavage site with a topoisomerase specific for that site, wherein the topoisomerase-DNA intermediate has one or more 5? single-strand tails; and (b) adding to the topoisomerase-DNA intermediate an acceptor RNA strand complementary to the 5? single-strand tail under conditions permitting a ligation of the covalently bound DNA strand of the topoisomerase-DNA intermediate to the RNA acceptor strand and dissociation of the topoisomerase, thereby covalently joining the DNA strand to the RNA strand. The present invention also provides a method of tagging a 5? end of an RNA molecule. The present invention further provides a DNA-RNA molecule which has been joined in vitro by the use of a topoisomerase. The present invention also provides a method of tagging a 5? end of an mRNA.

Abstract: A method of diagnosing a disease that includes obtaining experimental data on gene selections. The gene selection functions to characterize a cancer when the expression of that gene selection is compared to the identical selection from a noncancerous cell or a different type of cancer cell. The invention also includes a method of targeting at least one product of a gene that includes administration of a therapeutic agent. The invention also includes the use of a gene selection for diagnosing a cancer.

Abstract: A nucleotide chain to be modified, a nucleotide having a particular base that is different from bases constituting the nucleotide chain, an enzyme catalyzing addition of the nucleotide to the 3?-terminus of the nucleotide chain, a degrading enzyme acting specifically on the nucleotide, and a desired modifier for modifying the nucleotide chain are allowed to coexist in a buffer solution as a mixture solution such that: the nucleotide is added to the 3?-terminus of the nucleotide chain; the sequence of the added nucleotide is degraded to form, at the 3?-terminus of the nucleotide chain, a functional group capable of binding to the modifier; and the 3?-terminus of the nucleotide chain having the functional group thus formed is directly modified with the modifier. The reactions at three stages continuously proceed in the mixture solution. As a result, simplified procedures and reduced reaction time can be achieved.

Abstract: A method of synthesizing a cDNA chain using an insoluble carrier having on the surface thereof a polymer substance containing a first unit having a group derived from a phosphoric ester composing the hydrophilic portion of phospholipid, and a second unit having a group derived from carboxylic acid having an electron-attractive substituent bound to a carbonyl group, which includes immobilizing a polynucleotide for DNA elongation; bringing a solution containing an RNA fragment, nucleotide monomers, and a reverse transcriptase or an enzyme having polymerase activity into contact with the surface of the insoluble carrier; and allowing the polynucleotide for DNA elongation immobilized on the surface of the carrier to elongate using the RNA fragment contained in the solution as a template, to thereby form a single-strand cDNA.

Abstract: The present invention is directed to the thermostable DNA-polymerase protein of the archaeal ampullavirus ABV (Acidianus Bottle-shaped virus) and the nucleic acid encoding said DNA polymerase. The invention also relates to method of synthesizing, amplifying or sequencing nucleic acid implementing said DNA polymerase protein and kit or apparatus comprising said DNA polymerase protein.

Abstract: Methods, reagents, and kits for (mis)ligating oligonucleotide probes or for identifying at least one target nucleotide are disclosed. One can enhance the generation of misligation product using a ligase under reaction conditions and with reagents where that particular ligase is prone to misligation. Alternatively, one can decrease or avoid generating misligation products using a particular ligase under reaction conditions and using reagents where that ligase is at least less prone to misligation. In certain embodiments, the recombinant ligase from Archaeoglobus fulgidus (Afu) is employed due to its unique misligation properties.

Abstract: Compositions and methods are disclosed for automated storing, tracking, retrieving and analyzing biological samples, including dry storage at ambient temperatures of nucleic acids, proteins (including enzymes), and cells using a dissolvable dry storage matrix that permits recovery of biologically active materials. RFID-tagged biological sample storage devices featuring dissolvable or dissociable matrices are described for use as supports of biological samples, which matrices can be dried and subsequently rehydrated for sample recovery. Also disclosed are computer-implemented systems and methods for managing sample data.

Abstract: The present invention provides a nucleic acid synthesis method capable of continuously carrying out an extension reaction and a single molecule sequencing method capable of obtaining base information accurately at high speed. A method for synthesizing a nucleic acid, including the steps of: forming a complex of a target nucleic acid hybridized to a primer oligonucleotide and a DNA polymerase ?; allowing the DNA polymerase ? to incorporate a fluorescently-labeled dNTP so that the fluorescently-labeled dNTP is bound to the 3? end of the primer oligonucleotide; and allowing the DNA polymerase ? to continuously incorporate fluorescently-labeled dNTP to extend a nucleic acid complementary to the target nucleic acid from the 3? end of the bound fluorescently-labeled dNTP.

Abstract: A novel artificial nucleic acid base pair which is obtained by forming a selective base pair by introducing a group having steric hindrance (preferably a group having steric hindrance and static repulsion and a stacking effect) and can be recognized by a polymerase such as DNA polymerase; a novel artificial gene; and a method of designing nucleic acid bases so as to form a selective base pair with the use of steric hindrance, static repulsion and stacking effect at the base moiety of the nucleic acid. An artificial nucleic acid comprising these bases; a process for producing the same; a codon containing the same; a nucleic acid molecule containing the same; a process for producing a non-natural gene by using the same; a process for producing a novel protein by using the above nucleic acid molecule or non-natural gene, and the like.

Abstract: Disclosed are compositions and a method for of amplifying nucleic acid sequences useful for detecting the presence of molecules of interest. The method is useful for detecting specific nucleic acids in a sample with high specificity and sensitivity. The method also has an inherently low level of background signal. A preferred form of the method consists of a DNA ligation operation, an amplification operation, and a detection operation. The DNA ligation operation circularizes a specially designed nucleic acid probe molecule. This operation is dependent on hybridization of the probe to a target sequence and forms circular probe molecules in proportion to the amount of target sequence present in a sample. The amplification operation is rolling circle replication of the circularized probe. A single round of amplification using rolling circle replication results in a large amplification of the circularized probe sequences.

Abstract: Methods for measuring environmental parameters using chemical recording are provided. In some embodiments, the methods include generating a polymer comprising an ordered series of chemical units, wherein the position and number of each chemical unit in the polymer is indicative of a reading of the environmental state variable at a given point in time. The presently disclosed subject matter also provides compositions that can be employed in and/or that employ the disclosed methods for recording environmental state variables.

Abstract: The present invention discloses the engineering of a human enzyme with arginine hydrolytic activity suitable for human therapy. An enzyme comprising of a human sequence is not likely to induce adverse immunological responses and thus is expected to constitute a superior therapeutic. Since the human genome does not encode arginases with the proper high affinity catalytic properties (i.e., for example, a low Km and high catalytic activity, kcat) an appropriate arginase can be engineered by modifying an enzyme with related catalytic activity. For example, the human enzyme PAD4 can hydrolyze arginine in peptide substrates but does not have activity for free arginine. First, a high throughput assay was developed for detecting arginine activity by monitoring the formation of the hydrolytic product citrulline. Then, using a combination of rational design and iterative mutation and screening PAD4 mutants were identified and isolated exhibiting high affinity free arginine metabolic activity.

Abstract: A method of amplifying a nucleic acid is provided which comprises: generating a first nucleic acid primer from a first nucleic acid sequence; combining the first nucleic acid primer with a first polymerase and a first circular nucleic acid probe, wherein the first circular nucleic acid probe contains at least one antisense sequence to a second nucleic acid sequence and at least one antisense sequence to the first nucleic acid primer; producing at least one repeat of a sequence copy of the first circular nucleic acid probe by rolling circle amplification using the first polymerase, wherein the sequence copy contains at least the second nucleic acid sequence; generating a second nucleic acid primer from the second nucleic acid sequence; combining the second nucleic acid primer with a second polymerase and a second circular nucleic acid probe, where the second circular nucleic acid probe contains at least one antisense sequence to the second nucleic acid primer; and producing at least one repeat of a sequence co

Abstract: The invention provides methods for controlling the density of different molecular species on the surface of a solid support. A first mixture of different molecular species is attached to a solid support under conditions to attach each species at a desired density, thereby producing a derivatized support having attached capture molecules. The derivatized support is treated with a second mixture of different molecular species, wherein different molecular species in the second mixture bind specifically to the different capture molecules attached to the solid support. One or more of the capture molecules can be reversibly modified such that the capture molecules have a different activity before and after the second mixture of molecular species are attached. In particular embodiments, the different molecular species are nucleic acids that are reversibly modified to have different activity in an amplification reaction.

Abstract: A method for generating human IgG1 antibodies with enhanced Fc effector function is disclosed. In practicing the method, an IgG1 Fc look-through mutagenesis (LTM) coding library directed at four receptor-contact regions of the Fc CH2 portion of in human IgG1 Fc is expressed in a system in which the mutated Fc fragments are displayed on the surfaces of the expression cells. The fragments are then screened for altered binding affinity to a selected Fc receptor or other Fc-binding protein. The selected mutations may be used, in turn, to guide the selection of multiple substitutions in the construction of a walk-through mutation (WTM) library, for generating additional Fc fragment mutations with desired binding properties. The antibodies so produced have a variety of therapeutic and diagnostic applications.

Abstract: The present invention provides thermostable enzymes, such as DNA polymerases and restriction endonucleases, that are substantially free from contamination with nucleic acids. The invention also provides methods for the production of these enzymes, and kits comprising these enzymes which may be used in amplifying or sequencing nucleic acid molecules, including through use of the polymerase chain reaction (PCR).

Abstract: The present invention relates to methods and reagents for detecting analytes, e.g. nucleic acids. The new methods and reagents allow a simple and sensitive detection even in complex biological samples.

Abstract: A primer set for amplifying a region including a site to be detected of CT2C9*3 in the C-YT2C9 gene by a gene amplification method is provided, wherein the primer set can amplify the region specifically. A pair of primer set is used including a forward primer consisting of the base sequence of SEQ ID NO: 4 as well as a reverse primer consisting of the base sequence of SEQ ID NO: 17. The use of this primer set makes it possible to specifically and efficiently amplify a target region including the site where a polymorphism, CYP2C9*3, of the CYP2C9 gene is generated.

Abstract: The present invention describes primers, methods and kits for amplifying and identifying HLA alleles. Using these primers, all HLA alleles at a single locus can be amplified using either a multiplex or non-multiplex PCR approach. Within sets of the primers, control primer pairs may be used to produce control amplicons of a predetermined size from an HLA allele only if a particular HLA locus is present in the sample. The present invention further describes primers for sequencing HLA alleles following amplification. Methods and kits for using the primers are also disclosed.

Abstract: An embodiment of a system for reducing crosstalk in a parallel sequencing platform is described that comprises a substrate with a plurality of individual reaction environments that include a species of nucleic acid template, and a plurality of spatially localized reactants, wherein the localized reactants minimize the transmission of reaction products to a neighboring reaction environment due to a relative position of the localized reactants in the reaction environment.

Abstract: One embodiment of the invention is a method of producing an oligonucleotide extended by a single nucleotide base. An oligonucleotide and an extension terminating nucleotide are mixed with an enzyme having terminal transferase activity. The reaction produces an oligonucleotide extended by a single base. The extended oligonucleotide may be used as a size standard for single base extension reactions. Another embodiment of the invention is a method of producing a mixture of oligonucleotides extended by different single bases. An oligonucleotide, a first extension terminating nucleotide, and a second extension terminating nucleotide are mixed with an enzyme having terminal transferase activity. The first and second extension terminating nucleotides comprise different nucleotide bases and are labeled with different labels. The identity of the different extension terminating nucleotides (and hence the extended oligonucleotides) may be ascertained by reference to the specific label incorporated.

Abstract: The present invention provides methods and kits useful for enriching, identifying and quantifying methylated DNA3 particularly hypermethylated CpG islands by digesting a sample with a methylation-sensitive restriction endonuclease and capturing methylated restriction fragments with a methyl-binding capture reagent. The methods of the invention may be used in the detection of cancer, particularly detection of prostate cancer.

Abstract: Methods and systems for removing masking agents from test samples, e.g., DNA-containing samples obtained from living subjects, when they are submitted for or subjected to molecular assays. The present invention allows molecular assays of nucleic acids in bodily fluids and excretions, such as urine, blood, blood serum, amniotic fluid, spinal fluid, conjunctival fluid, salivary fluid, vaginal fluid, stool, seminal fluid, and sweat to be carried out with greater sensitivity. The masking agents are suppressed by contacting a test sample with an amount of one or more divalent metal chelators and an amount of one or more chelator enhancing components. The amounts of the divalent metal chelator(s) and the chelator enhancing component(s) are selected such that interference of a masking agent on a molecular assay of a nucleic acid-containing test sample are suppressed, and upon contact with the divalent metal chelator(s)/chelator enhancing component(s), the masking agents are suppressed.

Abstract: A process and apparatus for DNA sequencing is provided. In the field of DNA analysis, an iterative process is disclosed wherein an apparatus with a set of recognition chambers in which a species of recognition element nucleotides are differentially added and subjected to a polymerization reaction allows recognition of which species is next in sequence on a template strand as measured by a detector in a detection area. The position in the sequence is then completed by addition of a saturating amount of building element to complete the polymerization reaction on all structure strands. The process is repeated until the sequence is identified.

Abstract: Subject matter of the invention is a method for amplification of nucleic acids, wherein a successful binding of a primer and a facilitator onto the template is required for amplification. The invention further comprises the inventive facilitator, inventive kits comprising a facilitator and the use of said inventive method, facilitator or kit.

Abstract: A biological sample reaction chip, including: a plurality of reaction vessels; a first channel connected to one end of each of the reaction vessels and comprising an opening for introducing a reaction solution; and a second channel connected to the other end of each of the reaction vessels, wherein when a capillary force of the first channel is defined as A, while a capillary force of connected portions between the reaction vessels and the first channel as B, a capillary force of the reaction vessels as C, a capillary force of connected portions of the second channel and the reaction vessels as D, and a capillary force of the second channel as E, the following is established: A<B<C<D and E<D.

Abstract: Methods are provided for multiplexed amplification of selected targets and analysis of the amplified targets. In preferred aspects the amplification and analysis take place on the same solid support and preferably in a localized area such as a bead or a feature of an array. In preferred aspects the analysis is a determination of sequence at one or more locations in the amplified target. The methods may be used for genotyping, sequencing and analysis of copy number.

Abstract: We describe a method of selecting an enzyme having replicase activity, the method comprising the steps of: (a) providing a pool of nucleic acids comprising members each encoding a replicase or a variant of the replicase; (b) subdividing the pool of nucleic acids into compartments, such that each compartment comprises a nucleic acid member of the pool together with the replicase or variant encoded by the nucleic acid member; (c) allowing nucleic acid replication to occur; and (d) detecting amplification of the nucleic acid member by the replicase. Methods for selecting agents capable of modulating replicase activity, and for selecting interacting polypeptides are also disclosed.

Abstract: Provided herein are methods and agents for ligation-based exponential ribonucleic acid amplification followed by detection using a nucleic acid polymerization reaction employing terminal-phosphate-labeled nucleotides including three or more phosphates as substrates for nucleic acid polymerase.

Abstract: Provided are novel nucleotides, nucleoside, and their derivatives described herein, that can be used in DNA sequencing technology and other types of DNA analysis. In one embodiment, the nucleotide or nucleoside with an unprotected 3?-OH group is derivatized at the nucleobase to include a fluorescent dye attached via a linker to a photocleavable terminating group. The photocleavable-fluorescent group is designed to terminate DNA synthesis as well as be cleaved so that DNA oligomers can be sequenced efficiently in a parallel format. The design of such rapidly cleavable fluorescent groups on nucleotides and nucleosides can enhance the speed and accuracy of sequencing of large oligomers of DNA in parallel, to allow rapid whole genome sequencing, and the identification of polymorphisms and other valuable genetic information, as well as allowing further manipulation and analysis of nucleic acid molecules in their native state following cleavage of the fluorescent group.

Abstract: Cellular mRNA-protein (mRNP) complexes are partitioned in vivo by contacting a biological sample with at least one ligand that specifically binds at least one component of a mRNP complex. Suitable biological samples comprise at least one mRNA-protein (mRNP) complex and include cell cultures, cell extracts, and whole tissue, including tumor tissue. Ligands include antibodies that specifically bind RNA-binding or RNA-associated proteins present in the mRNP complex. The mRNP complex is separated by binding the ligand with a binding molecule specific for the ligand, where the binding molecule is attached to a solid support. The mRNP complex is collected by removing the mRNP complex from the solid support. After collecting the mRNP complex, the mRNA bound within the complex may be characterized and identified. Subsets of the total mRNA population of a cell may accordingly be characterized, and a gene expression profile of the cell obtained.

Abstract: The invention relates to the field of polynucleotide amplification. More particularly, the invention provides methods, compositions and kits for amplification of (i.e., making multiple copies of) a multiplicity of different polynucleotide template sequences using a randomly primed RNA/DNA composite primer.

Abstract: The invention relates to compounds including at least one pentopyranosyl nucleic acid and a biomolecule conjugated through a covalent linkage to the at least one pentopyranosyl nucleic acid. The pentopyranosyl nucleic acid has at least two pentopyranosyl nucleotide subunits covalently linked between carbon 4 and carbon 2 of their respective pentopyranosyl rings. The pentopyranosyl nucleic acid does not non-covalently bond or hybridize to naturally occurring DNA or RNA. The invention also relates to compounds comprising a biotin molecule and a pentopyranosyl nucleic acid conjugated through a covalent linkage to the biotin molecule. Processes for preparing conjugates are also described.

Abstract: A method for analysing a target polynucleotide having distinct units of nucleic acid sequence comprising: (i) forming a first polynucleotide which is a concatemer having multiple repeating target polynucleotide sequences; (ii) forming on the first polynucleotide a second polynucleotide hybridised to a portion of one or more of the target polynucleotides, such that the portion hybridised, or the portion not hybridised, corresponds to a sequence unit on the target, and determining the sequence unit on the target.

Abstract: The present invention provides a series of artificial CpG-containing single-stranded oligodeoxynucleotides (ODNs), each of which is consisted of single-stranded oligodeoxynucleotide DNA molecule containing one or more CpG(s), wherein said ODNs can stimulate human peripheral blood mononuclear cell (PBMC) to produce antiviral substances. These ODNs can protect the cells against the attack from virus, wherein said virus is preferably selected from the group consisted of influenza virus and single-stranded positive strand RNA virus such as SARS virus, hepatitis C virus, dengue virus and Japanese encephalitis virus. Moreover, the antiviral use of artificial CpG ODNs and its use for treating and preventing viral infection are also provided.

Abstract: A construct in which at least a part of the magnetic material is coated with a polyhydroxyalkanoate (PHA), and a method for producing a construct by immobilizing a PHA synthesizing enzyme on the surface of the magnetic material, thereby biosynthesizing and coating a PHA.

Abstract: Methods and kits for use in isolating, labeling or detecting a small polynucleotide of interest from a sample. The method entails hybridizing the small polynucleotide to a capture probe, lengthening the small polynucleotide by primer extension and/or ligation, and degrading the capture probe to provide a single stranded extension product. The kits include the capture probe and additional reagents for the extension, ligation and/or degradation reactions.

Abstract: Biological samples containing nucleic acids, RNA and DNA, are freed from bound proteins by incubation with a chaotropic agent such as a guanidium salt, and the mixture is readied for further processing by dilution of such agent to a concentration below 0.05 M without physical isolation of RNA and DNA from one another or from other components of the reaction mixture. Methods include such preparation and further processing, such as amplification and detection, which may be performed in a single container. Chaotropic agent may be supplied as a dried reagent adhered to a container. Kits may include such reagents, alone or with amplification reagents.

Abstract: The invention provides an improved cell free amplification method capable of producing large quantities of therapeutic-quality nucleic acids and methods of using the synthesized nucleic acid in research, therapeutic and other applications—The methods combine several different state-of-the-art procedures and coordinate their applications to affordably synthesize nucleic acids for therapeutic purposes. It combines in vitro rolling circle amplification, high fidelity polymerases, high affinity primers, and streamlined template specifically designed for particular applications. For expression purposes, the templates contain an expression cassette including a eukaryotic promoter, the coding sequence for the gene of interest, and a eukaryotic termination sequence.

Abstract: Methods of preparing nucleic acid from polysaccharide-containing samples for detection by providing one or more glycosidases to the sample to degrade polysaccharides are provided. The nucleic acids can further be extracted from the sample. The method is particularly useful for detecting nucleic acid in samples with high starch content.

Abstract: The present invention relates to a method for breast cancer diagnosis/prognosis comprising the following steps: A—extracting the nucleic material from a biological sample; B—using at least one pair of amplification primers to obtain amplicons of at least one target sequence of the nucleic material; C—using at least one detection probe to detect the presence of said amplicons, characterized in that, during step B, said pair of primers comprises at least one amplification primer comprising at least 10 nucleotide motifs of a nucleotide sequence selected from SEQ ID No. 1 to SEQ ID No. 8 and/or, during step C, said detection probe comprises at least 10 nucleotide motifs of a nucleotide sequence selected from SEQ ID No. 9 and SEQ ID No. 10. The invention also relates to amplification primers and hybridization probes which can be used in said method, and to a kit for the diagnosis/prognosis of breast cancer.

Abstract: Novel linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye are provided. These linkers faciliate the efficient transfer of energy between a donor and acceptor dye in an energy transfer dye. One of these linkers for linking a donor dye to an acceptor dye in an energy transfer fluorescent dye has the general structure R21Z1C(O)R22R28 where R21 is a C1-5 alkyl attached to the donor dye, C(O) is a carbonyl group, Z1 is either NH, sulfur or oxygen, R22 is a substituent which includes an alkene, diene, alkyne, a five and six membered ring having at least one unsaturated bond or a fused ring structure which is attached to the carbonyl carbon, and R28 includes a functional group which attaches the linker to the acceptor dye.

Abstract: A method for DNA reassembly after random fragmentation, and its application to mutagenesis of nucleic acid sequences by in vitro or in vivo recombination is described. In particular, a method for the production of nucleic acid fragments or polynucleotides encoding mutant proteins is described. The present invention also relates to a method of repeated cycles of mutagenesis, shuffling and selection which allow for the directed molecular evolution in vitro or in vivo of proteins.

Abstract: The present invention provides improved techniques and reagents for producing nucleic acid molecules. In certain preferred embodiments, the nucleic acid molecules are modular vectors. In certain preferred embodiments, the nucleic acid molecules are produced in polymerase chain reactions employing terminator primer residues.

Abstract: Provided are previously uncharacterised enhancer elements from the preprotachykinin-A (PPTA) which shows transcriptional enhancement activity in Substance P (SP) expressing cells. These are termed ECR1 and ECR2. The invention provides nucleic acids comprising these sequences and variants or fragments thereof, plus methods and materials based thereon, such as transformed host cells, transgenic animals, and screening and expression systems.

Abstract: The invention relates to the generation and characterization of stable MMLV reverse transcriptase mutants. The invention also discloses methods of using stable MMLV reverse transcriptase mutants.

Abstract: The invention relates to compositions, kits, and methods for detecting, characterizing, preventing, and treating prostate cancer. FKBP markers are provided, wherein changes in the levels of expression of one or more of the FKBP markers is correlated with the presence of prostate cancer.

Abstract: The inventions provides a method for identifying a target virus in an infected subject comprising the steps of designing a pair of degenerate primers corresponding to highly conserved regions of the target virus; designing a pair of species-specific primers according to highly variable sequences within the conserved regions of the target virus; preparing the species-specific probes according to the larger sequence variations within the conserved regions of the target virus, which are amplified with the species-specific primers as obtained; preparing a test sample by amplifying total nucleic acid of the infected subject with the degenerate primers as obtained; contacting the test sample with the species-specific probes as obtained; and detecting a hybridization between the species-specific probe and the test sample, wherein the hybridization indicates the target virus is identified in the infected subject. The primers and probes for detecting a garget virus are also provided.