[go: up one dir, main page]

WO2002040663A1 - Acides nucleiques permettant de detecter des champignons et procede de detection de champignons a l'aide desdits acides nucleiques - Google Patents

Acides nucleiques permettant de detecter des champignons et procede de detection de champignons a l'aide desdits acides nucleiques Download PDF

Info

Publication number
WO2002040663A1
WO2002040663A1 PCT/JP2001/009939 JP0109939W WO0240663A1 WO 2002040663 A1 WO2002040663 A1 WO 2002040663A1 JP 0109939 W JP0109939 W JP 0109939W WO 0240663 A1 WO0240663 A1 WO 0240663A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
acid mixture
fungus
detecting
candida
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/JP2001/009939
Other languages
English (en)
Japanese (ja)
Inventor
Koji Yokoyama
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SSP Co Ltd
Original Assignee
SSP Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SSP Co Ltd filed Critical SSP Co Ltd
Publication of WO2002040663A1 publication Critical patent/WO2002040663A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae

Definitions

  • the present invention relates to a nucleic acid mixture for detecting a fungus and a method for detecting a fungus using the mixture.
  • the main pathogens of deep mycosis are Candida (hereinafter abbreviated as G.) genus fungi, Aspergillus (hereinafter abbreviated as A.) genus fungi, and Cryptococcus (abbreviated as Gr. Below). ) Genus fungi are known.
  • Major pathogens of the genus Candida include Candida albicans (G. albicans), Candida albicans 'G. glabrata', Candida albicans 'G. tropical is', and Candida albicans (G. parapsi losis) Candida albicans 'G. dubl iniensis and other major pathogens of the genus Cryptococcus include Cryptococcus neoformans (Gr.
  • Neoformans The main pathogens of the genus Aspergillus include A. fumigatus, A. flavus, A. niger, and the two gar groups, Aspergillus drans (A. n). idurans) and Aspergillus' terreus (A. terreus), and it is desired to rapidly detect, classify, and identify these strains.
  • An object of the present invention is to provide a nucleic acid used for detecting fungi causing deep mycosis, particularly fungi of the genus Candida and Cryptococcus, and to use the nucleic acid in a simple, rapid, specific and specific manner. It is an object of the present invention to provide a highly sensitive method for detecting fungi of deep mycosis, particularly fungi of the genus Candida and cryptococci.
  • the present inventors have conducted various studies on the genes of various fungal fungi, especially Candida fungi and Cryptococcus fungi, and as a result, the fungi causing these various fungal diseases have been identified.
  • a part of the cytochrome b gene present in mitochondria was amplified and its sequence was successfully decoded.
  • the present inventors found specific sequences for each type of Candida fungus, and amplified a part of the cytochrome b gene of each species based on them.
  • a universal primer mixture has been designed to be able to do so.
  • the present invention relates to the present invention, wherein each of a plurality of base sequences represented by SEQ ID NOS: 1, 2, 3 or 4, each showing a plurality of base sequences (provided that thymine at any position is substituted with peracil. Or at least two or more of its complementary base sequences, each consisting of at least 15 bases continuous from the 3 ′ end, or hybridizing with each base sequence under stringent conditions.
  • nucleic acid mixture for detecting a fungus comprising a mixture of at least two or more nucleic acids having a base sequence.
  • the present invention relates to at least a plurality of base sequences represented by SEQ ID NOS: 1 and 2 in the sequence listing (though thymine at any position may be substituted with peracyl).
  • a base sequence consisting of at least 15 bases continuous from the 3 ′ end or a mixture of the base sequence and a nucleic acid having a base sequence that hybridizes under stringent conditions.
  • a primer on one side and has at least two or more of a plurality of base sequences represented by SEQ ID NO: 3 or 4 in the sequence listing (though thymine at any position may be substituted with peracyl).
  • SEQ ID NO: 3 or 4 in the sequence listing (though thymine at any position may be substituted with peracyl).
  • 'A nucleic acid amplification method using a nucleotide sequence comprising at least 15 nucleotides continuous from the terminal or a mixture of nucleic acids having a nucleotide sequence that hybridizes with the nucleotide sequence under stringent conditions as a reverse primer.
  • a method for detecting a fungus comprising amplifying a test nucleic acid and detecting an amplification product.
  • the present invention provides a method for identifying a fungus, comprising determining the base sequence of the amplification product.
  • the present invention further provides a kit for detecting a fungus containing the nucleic acid of the present invention.
  • the present invention provides a fungal detection probe obtained by labeling the nucleic acid mixture of the present invention.
  • the present invention provides a capture probe for detecting a fungus, wherein the nucleic acid mixture of the present invention is bound to a carrier.
  • the present invention provides the use of the nucleic acid mixture of the present invention for producing a probe or primer for detecting a fungus.
  • a fungal nucleic acid for use in detecting various fungi and a method for detecting a fungus using the nucleic acid are provided.
  • the nucleic acid mixture of the present invention can be used as a primer for an amplification reaction or as a probe for direct detection.
  • the nucleic acid mixture of the present invention as a primer for an amplification reaction, it has become possible to detect a plurality of fungi having different nucleotide sequences by a single amplification reaction.
  • the sensitivity and specificity can be further increased by detecting the product amplified by the primer with a probe, and its clinical significance is great.
  • the nucleic acid mixture for detecting a fungus of the present invention comprises a plurality of base sequences each represented by SEQ ID NO: 1, 2, 3 or 4 (provided that thymine at any position is substituted with peracil. Or at least two of its complementary base sequences Among them, those having a base sequence consisting of at least 15 bases continuous from at least the 3 'end or a base sequence that hybridizes with the base sequence under stringent conditions.
  • the nucleic acids of the present invention may be DNA or RNA.
  • nucleic acid mixture of the present invention at least 3 ′ of a plurality of base sequences represented by SEQ ID NOs: 1, 2, 3 or 4 (provided that thymine at any position may be substituted with peracyl)
  • a nucleotide sequence consisting of at least 15 bases continuous from the terminal is preferable, and a nucleic acid mixture represented by SEQ ID NO: 1, 2, 3, or 4 and having a plurality of nucleotide sequences is particularly preferable.
  • the nucleic acid mixture for detecting a fungus of the present invention comprises a nucleotide sequence represented by SEQ ID NO: 1, 2, 3 or 4 in the sequence listing (however, thymine at any position may be substituted with peracyl) or its complement.
  • a basic sequence consisting of at least 15 bases continuous from at least its 3 'end under stringent conditions the base sequence described above may be substituted with at least 3 bases. It may include deletions, insertions or additions.
  • “under stringent conditions” means that under the annealing conditions in ordinary PCR as described in the Examples below, the target nucleic acid hybridizes to type II and functions as a primer.
  • the buffer commonly used for PCR is PCR buffer (final concentration of 50 mM KG I, 10 mM Tris-HCl (pH 8.4-9. at 25 ° C), 1.5 mM MgG I 2 ) .Kit usually comes with a 10-fold concentration of X10 PGR buffer (or 10X PGR buffer), which is diluted.
  • nucleotide sequence represented by SEQ ID NO: 1, 2, 3 or 4 (however, thymine at any position may be substituted with peracyl) or at least the 3 '
  • a nucleic acid mixture having a base sequence consisting of at least 15 bases consecutive from the end and a sequence in which only one or two bases are substituted (excluding those in which the 3 'end is substituted) is usually used for the purpose of the present invention. Can be used for
  • the nucleic acid of the present invention can be easily produced by chemical synthesis using a commercially available DNA synthesizer or the like.
  • SEQ ID NOs: 1 to 4 indicate multiple bases with one letter, such as m, r, and w, respectively. Therefore, one sequence number indicates multiple sequences.
  • the nucleic acid of the present invention is a mixture of nucleic acids having at least two or more, and preferably all of these plural nucleotide sequences (including the above-mentioned portions and variants). By using such a mixture, various mutant strains of fungi can be detected.
  • the nucleic acid mixture of the present invention may comprise a region within the mitochondrial cytochrome b gene of a fungus, particularly a fungus belonging to the genus Candida, Cryptococcus, Aspergillus or Penicillium, preferably a fungus belonging to the genus Candida or Cryptococcus. It is what hybridizes.
  • Preferred fungal species that can be detected using the nucleic acid mixture of the present invention include Candida 'grabrata, Candida parapsilosis, Candida tropicalis, Candida' albicans, Cryptococcus' neoformans and Candida doubryensis But are not limited to these. However, SEQ ID NO: 3 (including the above-mentioned portions and variants) cannot be used for the detection of Candida albicans, Cryptococcus neoformans and Candida doubriniensis.
  • the mitochondrial cytochrome b gene is located in mitochondria, is difficult to recombine due to cytoplasmic inheritance, and is thought to undergo base changes in proportion to evolution. Therefore, it is suitable for detecting various fungi.
  • the region amplified when the nucleic acid combination of the present invention is used as a primer in a nucleic acid amplification method is a relatively variable portion, and it is easy to distinguish species, It has the advantage that it is easy to distinguish between DNA types within a species. Therefore, the species or strain of fungi can be identified by amplifying the test nucleic acid using the nucleic acid of the present invention as a primer and determining the base sequence of the amplification product.
  • the nucleic acid mixture of the present invention hybridizes with a region in the cytochrome b gene of the fungal mitochondria, so that the fungus can be detected by using the nucleic acid mixture of the present invention as a probe or a primer for a nucleic acid amplification method. it can.
  • the nucleic acid mixture of the present invention is used as a probe, it is preferable to use a nucleic acid mixture labeled with a labeling agent.
  • the labeled nucleic acid mixture of the invention is After the fungal gene in the test sample and the labeled probe are hybridized, the bound product of the fungal gene and the labeled probe or the unbound labeled probe is detected by an appropriate detection method suitable for the labeling agent.
  • Labeling agents used for probes are well known in this field, and include fluorescent labels, radioactive labels, enzyme labels, biotin and the like.
  • the hybridization of the sample, i.e., the fungal gene in the test sample, to the probe is performed by pretreating and purifying the sample by a usual method to obtain a test sample, and mixing it with a labeled nucleic acid mixture as a probe, and then room temperature. It can be carried out by treating at 10 to 70 ° C for 10 minutes to 48 hours.
  • the sample may be subjected to an amplification reaction in advance using the nucleic acid mixture of the present invention as a primer.
  • the nucleic acid mixture of the present invention is used as a primer for a nucleic acid amplification method
  • the nucleic acid mixture of the present invention is used as a primer to carry out an elongation reaction with a DNA polymerase or the like to amplify the gene, thereby obtaining a cytochrome of a Candida fungus and a Cryptococcus fungus.
  • a fungus can be detected by specifically amplifying only the b gene fragment and measuring the amplified product.
  • a labeled nucleic acid obtained by labeling the nucleic acid mixture of the present invention with a labeling agent as described above may be used as a primer.
  • the gene amplification method used herein include a PCR method, but are not particularly limited to this method. Any method using a short-chain oligonucleic acid as a primer to initiate gene synthesis is used. Can also be used.
  • the method for detecting a fungus of the present invention can be performed by detecting an amplification product amplified by the above method, ie, a cytochrome b gene fragment. Alternatively, it can also be carried out by fragmenting the amplified cytochrome b gene fragment with a restriction enzyme and comparing the generated patterns of the fragment.
  • the restriction enzymes used here are used alone or in combination.
  • Means for detecting a cytochrome b gene fragment amplified with an amplification product or a gene fragment further fragmented with a restriction enzyme is not particularly limited, and a normal gene detection method (for example, an electrophoresis method) can be used.
  • the amplification product is fractionated by, for example, electrophoresis and can be easily detected as a specific and sufficiently clear band for detection.
  • the primers are also used in the amplification reaction during the amplification reaction. By using a liponucleic acid mixture (ATP, UTP, GTP, CTP) as a raw material for DNA or RNA synthesis, amplification products can be directly detected with high sensitivity.
  • the amplified product can be directly detected with high sensitivity.
  • the labeling agent used for labeling is preferably a radioactive substance, a fluorescent substance, or the like. It can also be used as a primer for so-called real-time detection PCR, in which case the nucleic acid to be tested can be quantified. Therefore, “detection” as used herein includes quantitative detection.
  • a nucleic acid mixture having the nucleotide sequence shown in SEQ ID NO: 1 and SEQ ID NO: 2 (including the above-mentioned part and mutant thereof, the same applies hereinafter) is preferably used as a fore-side primer
  • the nucleic acid mixture having the nucleotide sequence shown in SEQ ID NO: 3 or SEQ ID NO: 4 is preferably used as a reverse primer. It is preferable to use a combination of these forward primer and linear primer, whereby a 396 bp region is amplified as specifically described in the Examples below.
  • the reagent kit for detecting a fungus of the present invention comprises the nucleic acid mixture of the present invention or a labeled nucleic acid obtained by labeling the nucleic acid mixture with an appropriate labeling agent.
  • the reagent kit of the present invention may contain the above-described nucleic acid, and may contain a labeling detection reagent, a buffer, and the like as other components.
  • the nucleic acid in the reagent kit of the present invention can be used as a primer or a probe. Whether a nucleic acid is used as a primer or a probe is different in only its means, and is essentially the same for detecting fungi.
  • the reagent kit is used as a reagent kit for detecting a fungus by the method using the above-described probe. Can be used as a reagent kit.
  • the reagent kit of the present invention when the nucleic acid mixture in the reagent kit is used as a probe May contain, in addition to the nucleic acid mixture for detecting Candida fungi and Cryptococcus fungi or the labeled nucleic acid mixture for detecting Candida fungi and Cryptococcus fungus of the present invention, a labeling detection reagent, a buffer, and the like. Good.
  • the reagent kit of the present invention is a nucleic acid mixture for detecting a Candida fungus and a Cryptococcus fungus of the present invention or a labeled Candida fungus and a Cryptococcus fungus detection.
  • nucleic acid synthases eg, DNA polymerase, RNA polymerase, reverse transcriptase, etc.
  • mixtures of deoxyribonucleotides dATP, dTTP, dGTP, dGTP
  • mixtures of ribonucleotides ATP, UTP
  • restriction enzymes e.g., restriction enzymes, buffers and the like.
  • kits include, for example, those obtained by adding the nucleic acid mixture of the present invention to commercially available PCR kits described in the following Examples.
  • the nucleic acid mixture of the present invention can be used as a capture probe by binding to a solid phase carrier.
  • the sandwich assay may be performed by combining two of the capture probe and the labeled probe.
  • the nucleic acid mixture is labeled with biotin, hybridized, and then captured with an avidin-bound carrier.
  • the sandwich assay if the nucleic acid mixture of the present invention is used for either one, specific measurement of the nucleic acid mixture of the present invention becomes possible, and there is no problem even if the specificity of the other nucleic acid mixture is slightly lower. .
  • the binding of the nucleic acid to the solid phase carrier can be easily performed by a well-known technique widely used in the field of DNA chips and the like.
  • nucleic acid mixtures 1 to 4 in the sequence listing were synthesized by chemical synthesis.
  • the various nucleic acid mixtures represented by SEQ ID NOs: 1 to 4 in the sequence listing are referred to as nucleic acid mixtures 1 to 4, respectively.
  • Example 2 (Candida albicans cytochrome b gene amplification reaction)
  • Candida albicans (Gandida albicans IF 40009) obtained by culturing in a port dextrose liquid medium were treated with 75% ethanol and sterilized.
  • the cells were obtained by centrifugation at 1500 g for 10 minutes, and about 40 ml of extraction buffer (0.9 M sorbitol, 10 mM EDTA, 10 mM Tris-HCl buffer, pH 7.1) was added, and the mixture was shaken well. Thereafter, the supernatant was discarded by centrifugation at 1500 g for 10 minutes, 30 ml of the same buffer was added.
  • extraction buffer 0.9 M sorbitol, 10 mM EDTA, 10 mM Tris-HCl buffer, pH 7.1
  • the supernatant was discarded after centrifugation, and 9 ml of the same buffer was added.
  • 1 ml of Zymolyase (Seikagaku Kogyo Co., Ltd. cell wall lysing enzyme, 10 mg / ml) dissolved in the same buffer, and the mixture was incubated at 37 ° C. for 1 hour and then treated with an ultrasonic washer for 1 minute. This was centrifuged at 1500 g for 10 minutes to obtain a supernatant, and further centrifuged at 20000 g for 15 minutes to obtain a precipitate. The precipitate was washed with an extraction buffer and centrifuged again at 20000 g for 15 minutes to obtain a mitochondrial fraction.
  • the total volume was made up to 50 I with water.
  • the reaction conditions were as follows.
  • the amplified DNA was separated and purified by agarose gel electrophoresis, and the nucleotide sequence was analyzed by the following method. Using a DNA sequencer (ABI Prism 377) manufactured by PerkinElmer and analyzed according to the Dye Terminator method according to the operation guide of the company. Nucleic acid mixture 2 was used on the forward side and nucleic acid mixture 4 was used on the reverse side as primers for base sequence analysis. The determined nucleotide sequence is shown in SEQ ID NO: 5 in the sequence listing.
  • Example 3 Candida 'glabrata cytochrome b gene amplification reaction
  • Example 2 In the same manner as in Example 2, using the nucleic acid mixture 1 and the nucleic acid mixture 3 obtained in Example 1, a part of the cytochrome b gene was amplified from Candida glabrata (Candida glabrata IFM 46843) by a PCR reaction, The nucleotide sequence was determined. Nucleic acid mixture 1 was used on the forward side and nucleic acid mixture 3 was used on the reverse side as primers for nucleotide sequence analysis. The nucleotide sequence is shown in SEQ ID NO: 6 in the sequence listing.
  • Example 4 (Amplification reaction of Candida parapsilosis cytochrome b gene) Using the nucleic acid mixture 1 and the nucleic acid mixture 3 obtained in Example 1 in the same manner as in Example 2, Candida parapsilosis was used. losis IFM 46829), a part of the cytochrome b gene was amplified by PCR, and its nucleotide sequence was determined. Nucleic acid mixture 1 was used on the forward side and nucleic acid mixture 3 was used on the reverse side as primers for nucleotide sequence analysis. The nucleotide sequence is shown as SEQ ID NO: 7 in the sequence listing.
  • Example 5 (Amplification reaction of Candida tropicalis cytochrome b gene)
  • Example 2 In the same manner as in Example 2, using the nucleic acid mixture 1 and the nucleic acid mixture 3 obtained in Example 1, a part of the cytochrome b gene from Candida tropical is (FM 46816) was subjected to a PCR reaction. It was amplified and its nucleotide sequence was determined. Base sequence solution Nucleic acid mixture 1 was used on the forward side and nucleic acid mixture 3 was used on the reverse side as the primer for analysis. The nucleotide sequence is shown in SEQ ID NO: 8 in the sequence listing.
  • Example 6 (Amplification reaction of Candida doubriniensis cytochrome b gene) Using the nucleic acid mixture 2 and nucleic acid mixture 4 obtained in Example 1 in the same manner as in Example 2, Candida doubriniensis (Candida doubriniensis) was used. A part of the cytochrome b gene from Candida dubl iniensis IFM 48313) was amplified by PCR, and its nucleotide sequence was determined. The nucleotide sequence analysis primer used was a nucleic acid mixture 2 on the forward side and a nucleic acid mixture 4 on the reverse side. The nucleotide sequence is shown in SEQ ID NO: 9 in the sequence listing.
  • Example II (Cryptococcus neoformans cytochrome b gene amplification reaction)
  • nucleic acid mixture 2 Using the nucleic acid mixture 2 and the nucleic acid mixture 4 obtained in Example 1 in the same manner as in Example 2, a portion of the cytochrome b gene is amplified from Cryptococcus neoformans (Gryptococcus neoformans IFM 46 138) by PCR. Then, its nucleotide sequence was determined. For the primer for nucleotide sequence analysis, the nucleic acid mixture 2 was used on the forward side, and the nucleic acid mixture 4 was used on the reverse side. The nucleotide sequence is shown as SEQ ID NO: 10 in the sequence listing.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Botany (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne des acides nucléiques utilisés pour détecter des champignons provoquant des infections fongiques qui entraînent une gêne profonde, notamment des champignons du genre Candida et Cryptococcus. Ces acides nucléiques sont des mélanges d'acides nucléiques permettant de détecter des champignons, qui comprennent au moins deux acides nucléiques sélectionnés dans le groupe constitué par des acides nucléiques à plusieurs séquences de base, chaque séquence contenant au moins 15 bases consécutives à partir de l'extrémité 3' d'au moins deux séquences de base représentée par SEQ ID NOS: 1, 2, 3 et 4.
PCT/JP2001/009939 2000-11-14 2001-11-14 Acides nucleiques permettant de detecter des champignons et procede de detection de champignons a l'aide desdits acides nucleiques Ceased WO2002040663A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2000-346887 2000-11-14
JP2000346887A JP2002142774A (ja) 2000-11-14 2000-11-14 真菌検出用核酸及びそれを用いた真菌の検出方法

Publications (1)

Publication Number Publication Date
WO2002040663A1 true WO2002040663A1 (fr) 2002-05-23

Family

ID=18820698

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2001/009939 Ceased WO2002040663A1 (fr) 2000-11-14 2001-11-14 Acides nucleiques permettant de detecter des champignons et procede de detection de champignons a l'aide desdits acides nucleiques

Country Status (2)

Country Link
JP (1) JP2002142774A (fr)
WO (1) WO2002040663A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116875721A (zh) * 2022-12-21 2023-10-13 广州医科大学附属第一医院 隐球菌的cfDNA在诊断隐球菌感染中的应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP4576489B2 (ja) * 2004-06-07 2010-11-10 国立大学法人 千葉大学 新規ポリヌクレオチド、それを用いた深在性輸入真菌症原因菌ペニシリウム・マルネッフェイ（Ｐｅｎｉｃｉｌｌｉｕｍｍａｒｎｅｆｆｅｉ）の検出用マーカ、プローブ、プライマー、検出方法およびキット

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0860503A1 (fr) * 1996-09-09 1998-08-26 SS Pharmaceutical Co., Ltd. Procede et materiau de detection de champignons
WO2000066773A2 (fr) * 1999-04-30 2000-11-09 Syngenta Limited Procedes

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0860503A1 (fr) * 1996-09-09 1998-08-26 SS Pharmaceutical Co., Ltd. Procede et materiau de detection de champignons
WO2000066773A2 (fr) * 1999-04-30 2000-11-09 Syngenta Limited Procedes

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
WANG L. ET AL.: "Mitochondrial cytochrome beta gene analysis of aspergillus fumigatus and related species", J. CLIN. MICROBIOL., vol. 38, no. 4, April 2000 (2000-04-01), pages 1352 - 1358, XP002908819 *
WANG L. ET AL.: "The identification and phylogenetic relationship of pathogenic species of aspergillus based on the mitochondrial cytochrome beta gene", MED. MYCOL., vol. 36, no. 3, 1998, pages 153 - 164, XP002908820 *
YOKOYAMA K. ET AL.: "Identification and phylogenetic relationship of the most common pathogenic candida species inferred from mitochondrial cytochrome beta gene sequences", J. CLIN. MICROBIOL., vol. 38, no. 12, December 2000 (2000-12-01), pages 4503 - 4510, XP002908818 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN116875721A (zh) * 2022-12-21 2023-10-13 广州医科大学附属第一医院 隐球菌的cfDNA在诊断隐球菌感染中的应用
CN116875721B (zh) * 2022-12-21 2024-03-19 广州医科大学附属第一医院 隐球菌的cfDNA在诊断隐球菌感染中的应用

Also Published As

Publication number Publication date
JP2002142774A (ja) 2002-05-21

Similar Documents

Publication Publication Date Title
JP2561053B2 (ja) ナイセリア・ゴノレエの検出方法、並びにそのための試薬およびキット
JPH10504973A (ja) 微生物検査室における日常的診断用の臨床検体からの通常の細菌病原体および抗生物質耐性遺伝子を迅速に検出および同定するための特異的および普遍的プローブおよび増幅プライマー
JP4662578B2 (ja) 抗生物質耐性微生物を同定するための方法およびキット
CA2623736C (fr) Compositions et methodes de detection d'une espece candidat
AU2003241562B2 (en) Molecular identification of Aspergillus species
US9434986B2 (en) Methods and kits used in the detection of fungus
AU727945B2 (en) Sequential hybridization of fungal cell DNA, and method for detecting fungal cells in clinical material
CN115803463A (zh) 用于诊断SARS-CoV-2的组合物、试剂盒以及通过使用所述试剂盒诊断SARS-CoV-2的方法
WO2002040663A1 (fr) Acides nucleiques permettant de detecter des champignons et procede de detection de champignons a l'aide desdits acides nucleiques
US20040235021A1 (en) Authentication of biologic materials using DNA-DNA hybridization on a solid support
WO1998010073A1 (fr) Procede et materiau de detection de champignons
JPH10210980A (ja) 乳酸菌検出用オリゴヌクレオチド及び該菌の検出方法
WO2002040713A1 (fr) Acides nucleiques permettant de detecter des champignons appartenant au genre candida et methode permettant de detection des champignons appartenant au genre candida a l'aide de ces acides nucleiques
JP5548389B2 (ja) ビソクラミス属に属する菌類の検出方法
JP2010004878A (ja) タラロマイセス属に属する菌類の検出方法
JP2010004879A (ja) ネオサルトリア属に属する菌類及びアスペルギルスフミガタスの検出方法
JP4576489B2 (ja) 新規ポリヌクレオチド、それを用いた深在性輸入真菌症原因菌ペニシリウム・マルネッフェイ（Ｐｅｎｉｃｉｌｌｉｕｍｍａｒｎｅｆｆｅｉ）の検出用マーカ、プローブ、プライマー、検出方法およびキット
JP2001299354A (ja) マイコバクテリウム属細菌検出用核酸
JP5697881B2 (ja) パエシロマイセスサトゥラタス及びパエシロマイセスディバリカタスの検出方法
JP2923778B1 (ja) 白紋羽病菌の特異的検出法
US20020172945A1 (en) Materials and methods for detection of pathogenic guignardia citricarpa
JP2006280273A (ja) 新規ポリヌクレオチド、それを用いた深在性輸入真菌症原因菌ヒストプラズマ属菌（Ｈｉｓｔｏｐｌａｓｍａ属菌には、Ｈｉｓｔｏｐｌａｓｍａｃａｐｓｕｌａｔｕｍｖａｒ．ｃａｐｓｕｌａｔｕｍ，Ｈｉｓｔｏｐｌａｓｍａｃａｐｓｕｌａｔｕｍｖａｒ．ｆａｒｃｉｍｉｎｏｓｕｍ，Ｈｉｓｔｏｐｌａｓｍａｃａｐｓｕｌａｔｕｍｖａｒ．ｄｕｂｏｉｓｉｉと有性世代の菌名であるＥｍｍｏｎｓｉｅｌｌａｃａｐｓｕｌａｔａを含む）の検出用マーカ、プローブ、プライマー、検出方法およびキット
HK40081979A (en) Composition for diagnosing sars-cov-2, kit, and method for diagnosing sars-cov-2 by using same
JPWO1998010073A1 (ja) 真菌類の検出用材料及び検出法
JPH11332596A (ja) シュードモナス・アエルギノーサの検出法及び検出用キット

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CA CN KR US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR

121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase