Ampdirect^® Gene Amplification

新型 PCR 扩增缓冲液

　　——无需抽提 DNA，直接 PCR 扩增！

　　Ampdirect^® Gene Amplification 是一款新开发的 PCR 缓冲液，可有效降低 PCR 反应体系里的蛋白质、糖类等各种 PCR 抑制物对 PCR 扩增的影响。应用广泛，适合对昆虫、植物、微生物、土壤、血液、石蜡切片等各类样品直接进行 PCR 扩增。

◆优势

●　省时省力省钱

　  无需 DNA 抽提，直接进行 PCR 反应，节省时间

●　适用于微量样品

　  因为不需要 DNA 抽提，不会产生样品损失，故此十分适用于微量样品

●　热启动酶

　  厂家自行开发的高性能聚合酶 BIOTAQ ™

●　支持后续实验操作

　  扩增产物可用于后续的测序和 RFLP 等片段分析

◆实验流程

◆原理

◆示例

注：动物样本，比如血液和黏膜细胞等可以直接加入 PCR 反应液，不需消化处理。固体样本，比如植物和动物组织需要在含有 SDS 和蛋白酶 K 的

       消化液中处理后再进行 PCR 反应，或者使用 FTA 卡收集植物组织中的 DNA，取带有 DNA 的 FTA 卡加入 PCR 反应液进行PCR 反应。

◆其他应用

● 血清中病毒检出

● 小鼠尾巴裂解液检测基因分型

● 植物、血液/ 滤纸血液、土壤

● 昆虫、石蜡切片等微量样品

   其他应用的详细内容请看相关资料！

相关资料

◆Ampdirect 推荐使用酶与不推荐使用酶

◆其他应用

Ampdiret 相关的FAQ

◆有关样品以及样品前处理

1．采集后长期保存的血液作为模板，能进行 PCR 吗？

实验证明，采集后冻存了长达五年时间的血或纸血（干燥血）作为模板，依然能进行PCR。必须长期稳定保存血液时，可使用 Whatman 公司的 FTA Card（核酸保存用卡）进行保存。关于血液（全血）对 PCR 的抑制作用，直接采集的血液（新鲜血）对 PCR 抑制作用更强。

另外，长久以来的经验证明，采用干燥血作为 PCR 模板时，对比用一般的实验纸保存，使用 FTACard 保存的血液进行 PCR 的结果更稳定，因此推荐使用 FTA Card 保存的血作为 PCR 模板。

2．从粪便中抽提的 DNA 模板，可用 PCR 检测出细菌吗？

粪便抽提的 DNA 对 PCR 有抑制作用，因此用稀释后的 DNA 反应。从粪便样品中简便进行 PCR 的应用例子，在“使用实时 PCR 法高灵敏度检测 Vero Toxin”中有介绍。将10%的粪便悬浊液热处理（95℃，5 分钟）后，将离心后的上清液作为 PCR 模板。实验证明，使用这个方法，大部分的粪便样品都能稳定进行 PCR 。

3．Ampdirect 可应用于 RT-PCR 吗？粪便里存在的 RNA 病毒也可以检测出来吗？

从粪便样品中简便检测 RNA 病毒的应用例子，可见“粪便中的 RNA 病毒检测”。将粪便悬浊液离心后的上清液，涂布在 Whatman 公司的 FTA Card（核酸保存用卡）上，干燥后，在卡上打（φ1.25 mm）就可作为模板使用。

Ampdirect Plus 可做为逆转录反应的反应液，逆转录酶选用的是 Invitrogen 公司的 M-MLVReverse Transcriptase（Cat No. 28025-013）。

4．样品的溶解处理最后步骤要在95℃，进行５分钟的热处理，这个热处理步骤是必须的吗？

因为要使溶解液中的 Proteinase K 失活，所以必须热处理。如果不进行热处理，Proteinase K 仍保持活性的话，在往 PCR 反应液里加入溶解液时，Proteinase K 会将 PCR 酶（Taq DNA Polymerase）降解掉，从而不能进行稳定的 PCR。实际上，这是从大部分得不到目的 PCR 产物的例子上取得的经验。所以，必须在最后对溶解液进行热处理（95℃，5 分钟）。

5．从植物的叶片抽提的 DNA 作为 PCR 模板进行反应，但得不到目的 PCR 产物。考虑到是植物里所含的糖类抑制了 PCR，使用 Ampdirect 可以改善这个情况吗？

至今为止，已有非常多客户反映，从实验植物、谷物、蔬菜、果蔬的叶片取样，用 Ampdirect 进行 PCR，都取得了良好的结果。不同植物对 PCR 的抑制作用也不尽相同，可参照“从植物取样的简便 PCR 实例”。在 PCR 前用溶解液进行前处理。如担心因为 PCR 被抑制而得不到目的 PCR 产物，用溶解处理液稀释５~10 倍，会对此情况有很大改善。

6．小鼠尾部用溶解液溶解了大约１个小时，还没完全溶解。可作为 PCR 的模板使用吗？

将小鼠尾巴１~５mm 浸入 100 μL 溶解液内，55℃下溶解约１小时后，如小鼠尾巴还未完全溶解，对 Proteinase K 进行失活处理（95℃，5 分钟）后，从溶解液中提取出来的 DNA 量（拷贝数）应该足以作为 PCR 模板，大部分情况下进行 PCR 没有问题。

与此对照，如果是将大条的小鼠尾巴（５ mm 以上）过夜处理以至完全溶解，溶解液会变成高粘度状态。这种情况下，过剩的 DNA 反而会抑制 PCR 反应，请用蒸馏水或 TE 将溶解液稀释 10 倍左右。请注意尽量不要让小鼠尾巴过度溶解。

另外，溶解处理后的溶解液不需离心，只需取上清液就可作为 PCR 模板使用。从以往经验得知，经溶解处理过的溶解液在冷藏保存条件下保存２~３年内皆可作为 PCR 模板使用。

7．不对唾液进行前处理就直接作为 PCR 模板是不可行的吧？“取样自口腔粘膜的 PCR”这个应用，是对口腔黏膜细胞用溶解液进行溶解处理，如果是用唾液的话，也必须用溶解液进行溶解处理吗？

目前，也有不少客户为了检测唾液中的微生物，不进行前处理，直接将唾液当成 PCR 模板使用。当作为模板进入 PCR 反应液的唾液中（数 μL）有大量微生物，可获得良好的 PCR 产物。与之相反，若微生物数量较少，根据唾液中的蛋白等物质对 PCR 抑制的不同程度，有可能得不到稳定的 PCR 产物。因此，推荐对溶液用溶解液进行溶解处理，再作为 PCR 模板。对唾液进行了溶解处理的话，可得到以下两种结果，①“抽提微生物的DNA”和②“降低唾液对 PCR 的抑制作用（分解唾液中的蛋白）”，也可得到稳定的 PCR 结果。

8．如何用 PCR 检测血浆、血清样品中的微生物？

血浆、血清这些全血，对PCR 有较强抑制作用，所以不能将全血直接用作 PCR 模板。因此，要用于“检测血清中的病毒”时，将血浆、血清用溶解液进行溶解处理，分解掉血浆、血清中的蛋白，降低对PCR 的抑制作用。

请注意，本方法适用于检测 DNA 病毒、细菌，不可用于检测 RNA 病毒。检测 RNA 病毒时，在溶解处理步骤，因为血浆、血清中存在 RNA 分解酶，从病毒外壳裸露出来的 RNA 就会被分解。

9、使用 FTA Card 和实验纸作为模板进行多样品 PCR 时，因为使用打孔器等工具可能会引起污染。有没有相应的对策呢？

采集血液样品时，首次打孔取样后，在下一次取样前，用打孔器在 FTA Card 或实验纸上没有血样的空白处空打３个孔，这样可保证打孔器上不会残留前一样品的模板（血样是白血球 DNA）。因为从客户处得知，“每次用打孔器取样时，用金伯利（Kimwipe）浸渍酒精擦拭打孔器的打孔部分，这个方法反而引起污染”，所以推荐空打孔的方法。但是，检测拷贝数多的 DNA，如线粒体 DNA 和质粒 DNA 等时，本方法不能防止污染。

参考文献：

●	Zoophilic feeding behaviour of phlebotomine sand flies in the endemic areas of cutaneous leishmaniasis of Sindh Province, Pakistan. Tiwananthagorn S, et al.
●	The sequestrate genus Rosbeeva T.Lebel & Orihara gen. nov. (Boletaceae) from Australasia and Japan: new species and new combinations. Lebel T, et al.
●	Tandem repeat inserts in 13S globulin subunits, the major allergenic storage protein of common buckwheat (Fagopyrum esculentum Moench) seeds. Khan N, et al.

Parasitol Res. 2012 Jan 14

・     Zoophilic feeding behaviour of phlebotomine sand flies in the endemic areas of cutaneous leishmaniasis of Sindh Province, Pakistan.

　    Tiwananthagorn S, et al.

样品：sand fly（沙蝇）、Leishmania（原虫）

Fungal Diversity (2012) 52: 49-71

・     The sequestrate genus Rosbeeva T.Lebel & Orihara gen. nov. (Boletaceae) from Australasia and Japan: new species and new combinations.

　    Lebel T, et al.

样品：菌類、FTA card        

Food Chemistry Volume 133, Issue 1, 1 July 2012, Pages 29-37

・     Tandem repeat inserts in 13S globulin subunits, the major allergenic storage protein of common buckwheat (Fagopyrum esculentum Moench) seeds.

　    Khan N, et al.

样品：荞麦（種子）

    

Acta Trop. 2012 Feb; 121(2): 93-8

・     Genotyping of sand fly species in Peruvian Andes where leishmaniasis is endemic.

　    Fujita M, et al.

样品：sand fly（沙蝇）

      

Zool J Linn Soc. 2012 Feb; 164(2): 304-27

・     Revision of the Euthalia phemius complex (Lepidoptera: Nymphalidae) based on morphology and molecular analyses.

　    Yago M, et al.

样品：昆虫（鱗翅目）

 

Limnology (3 November 2011), pp. 1-5

・     Surveillance of fish species composition using environmental DNA.

　    Minamoto T, et al.

样品：Environmental water

    

Am. J. Pot Res (2011) 88: 500-10

・     Characterization of Crossability in the Crosses between Solanum demissum and S. tuberosum, and the F1 and BC1 Progenies.

　    Sanetomo R, et al.

样品：土豆（種子） 

    

Systematic Botany (2011) 36(4): 836-44

・     A New Allotetraploid Species of Osmunda (Osmundaceae).

　    Tsutsumi C, et al.

样品：蕨类植物（根、叶） 

Am. J. Pot Res

・     Germplasm Release: Saikai 35, a Male and Female Fertile Breeding Line Carrying Solanum Phureja-Derived Cytoplasm and Potato Cyst Nematode Resistance (H1) and Potato Virus Y Resistance (Rychc) Genes.

　    Mori K, et al.

样品：土豆     

Neurochem Res. 2011 Nov;36(11):2127-35.

・     Ceruloplasmin protects against rotenone-induced oxidative stress and neurotoxicity.

　    Hineno A, et al.

样品：小鼠   

 

Bone. 2011 Nov;49(5):1027-36.

・    Insertional mutation in the Golgb1 gene is associated with osteochondrodysplasia and systemic edema in the OCD rat.

　    Katayama K, et al.

 

样品：大鼠  

  

Mycoscience

・    The genus Ponticulomyces (Physalacriaceae, Agaricales) from Japan.

　    Ushijima　S, et al.

样品：菌類、FTA card     

 

Trans R Soc Trop Med Hyg. 2011 Oct;105(10):561-7.

・    Leishmania species identification using FTA card sampling directly from patients’ cutaneous lesions in the state of Lara, Venezuela.

　     Kato H, et al.

样品：原虫（Leishmania）、FTA card  

 

Anticancer Res. 2011 Oct;31(10):3607-13.

・    Identification of The Distinctive Type i/XhoI+ Strain of Epstein-Barr Virus in Gastric Carcinoma in Peru.

　    Ordonez P, et al.

样品：EB病毒    

分析手法：PCR-RFLP法          

 

Mol Genet Metab. 2011 Sep 10.

・    Newborn screening for Pompe disease in Japan.

　    Oda E, et al.

样品：人体血液/滤纸血 

 

Blood. 2011 Sep 29.

・    Developmental origins and impact of BCR-ABL1 fusion and IKZF1 deletions in monozygotic twins with Ph+ acute lymphoblastic leukaemia.

　    Cazzaniga G, et al.

样品：滤纸血   

      

Mol Biol Evol. 2011 Sep 22.

・    Entangling Ancient Allotetraploidization in Asian Mitella: An Integrated Approach for Multilocus Combinations.

　    Okuyama Y, et al.

样品：植物 

  

Parasitol Res. 2011 Jul 8.

・    Prevalence of Trypanosoma sp. in cattle from Tanzania estimated by conventional PCR and loop-mediated isothermal of amplification (LAMP).

　    Laohasinnarong D, et al.

样品：牛血液、原虫（锥虫） 

 

J Agric Food Chem. 2011 Jul 13;59(13):6856-63.

・    Practicable group testing method to evaluate weight/weight GMO content in maize grains.

　    Mano J, et al.

样品：植物（玉米）

 

J Parasitol. 2011 Jun 14.

・    Molecular prevalence of different genotypes of theileria orientalis detected from cattle and water buffaloes in thailand.

　    Altangerel K, et al.

样品：牛、水牛血液、原虫 

 　

Euphytica

・    Reciprocal differences in DNA sequence and methylation status of the pollen DNA between F1 hybrids of Solanum tuberosum × S. demissum.

　    Sanetomo R, et al.

样品：植物花粉（土豆） 

 

J Vet Sci. 2011 Jun;12(2):191-3.

・    A simplified PCR assay for fast and easy mycoplasma mastitis screening in dairy cattle.

　    Higuchi H, et al.

样品：牛乳、細菌　 

 

Biol Pharm Bull. 2011;34(5):779-82.

・    Identification of dendrobium species used for herbal medicines based on ribosomal DNA internal transcribed spacer sequence.

　    Takamiya T, et al.

样品：植物  

 

Zootaxa 2905: 33-56 (3 Jun. 2011)

・    A survey of morphological variation in adult Meristogenys amoropalamus (Amphibia, Anura, Ranidae), with a description of a new cryptic species.

　    Shimada T, et al.

样品：青蛙

    

Plant Syst Evol (2011) 292: 177-188.

・    Phytogeographic aspects of Lysionotus pauciflorus sensu lato (Gesneriaceae) in the China, Japan and Taiwan regions: phylogenetic and morphological relationships and taxonomic consequences.

　    Kokubugata G, et al.

样品：植物  

 

FEMS Microbiol Ecol. 2011 Jan 11.

・    Microbial diversity with dominance of 16S rRNA gene sequences with high GC contents at 74℃ and 98℃ subsurface crude oil deposits in Japan.

　    Yamane K, et al.

样品：石油中采集的细菌   

 

J Mol Neurosci. 2011 Feb;43(2):217-24.

・    Decreased intake of sucrose solutions in orexin knockout mice.

　    Matsuo E, et al.

样品：小鼠   

             

Glycoconj J. 2010 Dec 21.

・    Excretion into feces of asialo GM1 in the murine digestive tract and Lactobacillus johnsonii exhibiting binding ability toward asialo GM1. A possible role of epithelial glycolipids in the discharge of intestinal bacteria.

　    Iwamori M, et al.

样品：細菌（Lactobacillus johnsonii、Lactobacillus casei）   

 

Mycologia. 2010 Dec 21.

・    Two species of Strobilomyces (Boletaceae, Boletales), S. seminudus and S. hongoi sp. nov. from Japan.

　    Sato H, et al.

样品：菌類  

 

Molecular Ecology Resources. 2010 Oct 28.

・    A primer set to determine sex in the small Indian mongoose, Herpestes auropunctatus.

　    Murata C, et al.

 样品：動物（猫鼬）

 

Entomological Science 2010 13: 303-310

・    An alien Sennertia mite (Acari: Chaetodactylidae) associated with an introduced Oriental bamboo-nesting large carpenter bee (Hymenoptera: Apidae: Xylocopa) invading the central Honshu Island, Japan.

　    Kawazoe K, et al.

样品：木匠蜂螨

  

Vector Borne Zoonotic Dis. 2010 Oct 18.

・    Natural Infections of Man-Biting Sand Flies by Leishmania and Trypanosoma Species in the Northern Peruvian Andes.

　    Kato H, et al.

样品：原虫（Leishmania and Trypanosoma Species） 

 

BMC Ecol. 2010 Oct 15;10:21.

・    Genetic structure of the oak wilt vector beetle Platypus quercivorus: inferences toward the process of damaged area expansion.

　    Shoda-Kagaya E, et al.

样品：昆虫  

 

Mycoscience. 2010 Oct 30.

・    Rapid detection for sporeless trait from Pleurotus pulmonarius culture extracts by using real-time PCR.

　    Okuda Y, et al.

样品：菌類  

分析手法：融解温度解析          

 

IBC 2010, vol.2, article no.12, pp.1-7

・    Genetic Quality Control of the Rat Strains at the National Bio Resource Project – Rat.

　    Kuramoto T, et al.

样品：大鼠、FTA card     

分析手法：Amp-FTA method             　

 

Int J Environ Res Public Health. 2010 Mar;7(3):814-26.

・    Molecular epidemiology for vector research on leishmaniasis.

　    Kato H, et al.

样品：昆虫、原虫    

分析手法：PCR-RFLP法           　

   

Am J Sports Med. 2010 Aug 19.

・    Cartilage Intermediate Layer Protein Gene Is Associated With Lumbar Disc Degeneration in Male, but Not Female, Collegiate Athletes.

　    Min SK, et al.

样品：人口腔粘膜細胞              

   

BMC Evol Biol. 2010 Jun 18;10:185.

・    Geographic variation in the damselfish-red alga cultivation mutualism in the Indo-West Pacific.

　    Hata H, et al.

样品：植物（紅藻）           

 

 

      

J Biochem. 2010 Aug 10.

・    NF-kappaB regulates the expression of Nucling, a novel apoptosis regulator, with involvement of proteasome and caspase for its degradation.

　    Tran NH, et al.

样品：小鼠           

 

Med Mycol. 2010 Jun;48(4):665-8.

・    Nourseothricin acetyltransferase: a new dominant selectable marker for the dermatophyte Trichophyton mentagrophytes.

　    Alshahni MM, et al.

样品：真菌           

 

J Plant Res. 2010 May 15.

・    Molecular database for classifying Shorea species (Dipterocarpaceae) and techniques for checking the legitimacy of timber and wood products.

　    Tsumura Y, et al.

样品：植物           

 

Forensic Toxicology 2010 July Volume 28, Number 2, 1-7

・    Chemical constituents and DNA sequence analysis of a psychotropic herbal product.

　    Kikuchi H, et al.

样品：植物           

 

J Clin Microbiol. 2010 Aug 18.

・    Use of FTA Cards for Direct Sampling of Patients' Lesions in the Ecological Study of Cutaneous Leishmaniasis.

　    Kato H, et al.

样品：原虫、FTA card        　

   

Vet Parasitol. 2010 Aug 4;171(3-4):207-15.

・    Generation of IFN-gamma-producing cells that recognize the major piroplasm surface protein in Theileria orientalis-infected bovines.

　    Yamaguchi T, et al.

样品：牛血液、原虫           　　

 

Jpn J Infect Dis. 2010 May;63(3):173-80.

・    Development of triplex SYBR green real-time PCR for detecting Mycoplasma pneumoniae, Chlamydophila pneumoniae, and Legionella spp. without extraction of DNA.

　    Kerdsin A, et al.

样品：    細菌 (Mycoplasma pneumoniae、Chlamydophila pneumoniae、Legionella spp.）

分析手法：    real-time PCR    

 

Am J Physiol Renal Physiol. 2010 Jun;298(6):F1341-50.

・    Phosphaturic action of fibroblast growth factor 23 in Npt2 null mice.

　    Tomoe Y, et al.

样品：小鼠           

 

Exp Eye Res. 2010 Jul;91(1):26-33.

・    A novel middle-wavelength opsin (M-opsin) null-mutation in the retinal cone dysfunction rat.

　    Xie B, et al.

样品：小鼠、FTA card        　

  

Am. J. Botany 2010 97: 373-387.

・    Developmental morphology of seedling and shoot and phylogenetic relationship of Diplobryum koyamae (Podostemaceae).

　    Koi S, et al.

样品：植物           

 

Food Control Volume 21, Issue 5, May 2010, Pages 599-605

・    Meat species identification based on the loop mediated isothermal amplification and electrochemical DNA sensor.

　    Ahmed MU, et al.

样品：食肉           

 

J Virol Methods. 2010 Feb;163(2):282-6.

・    Detection of noroviruses in fecal specimens by direct RT-PCR without RNA purification.

　    Nishimura N, et al.

样品：RNA病毒          

   

Theor Appl Genet. 2010 Jan;120(2):205-14.

・    DNA methylation in diploid inbred lines of potatoes and its possible role in the regulation of heterosis.

　    Nakamura S, et al.

 

样品：植物（potato）     

分析手法：RAPD法          

 

Genes Genet Syst. 2009 Oct;84(5):371-8.

・    Comparative differentiation in mitochondrial and chloroplast DNA among cultivated potatoes and closely related wild species.

　    Hosaka K, et al.

样品：植物（土豆）     

分析手法：PCR-RFLP法          

   

J Fish Dis. 2009 Oct;32(10):857-64.

・    Distribution of the introduced cyprinid herpesvirus 3 in a wild population of common carp, Cyprinus carpio L.

　    Uchii K, et al.

样品：魚類（鲤鱼）、DNA病毒        

     

Plant Physiol. 2009 Dec;151(4):2046-57.

・    The phytochrome-interacting factor PIF7 negatively regulates DREB1 expression under circadian control in Arabidopsis.

　    Kidokoro S, et al.

样品：植物（拟南芥）              

  

Divers. Distrib. 15 (6), 917-927 (2009)

・    Chloroplast DNA phylogeography of the endangered Japanese red maple (Acer pycnanthum): the spatial configuration of wetlands shapes genetic diversity.

　    Saeki I, et al.

样品：植物（枫树）           

 

Indian Journal of Science and Technology Vol.2 No. 10 (Oct 2009)

・    Development of activated sludge adapted to high concentrations of phenol and enhancement of its phenol removal ability by addition of a processed lignite.

　    Ohtsuki T, et al.

样品：活性汚泥        

分析手法：PCR-DGGE法         

 

Mol Ecol. 2009 Dec;18(23):4904-11.

・    Hybridization involving independent gametophytes in the Vandenboschia radicans complex (Hymenophyllaceae): a new perspective on the distribution of fern hybrids.

　    Ebihara A, et al.

样品：植物（蕨类）     

分析手法：PCR-SSCP法          

  

Appl Physiol Nutr Metab. 2009 Oct;34(5):926-32.

・    Is there a gender difference between ACE gene and race distance?

　    Min SK, et al.

样品：人 口腔粘膜細胞            

 

Int J Sports Med. 2009 Sep;30(9):691-4.

・    The cartilage intermediate layer protein gene is associated with lumbar disc degeneration in collegiate judokas.

　    Min SK, et al.

样品：人口腔粘膜細胞              

 

Biosci Biotechnol Biochem. 2009 Nov;73(11):2452-9.

・    Genome-wide identification, structure and expression studies, and mutant collection of 22 early nodulin-like protein genes in Arabidopsis.

　    Mashiguchi K, et al.

样品：植物（拟南芥）              

 

Plant Biotechnology 26(4), 435-441, 2009

・    Modification of the surface carbohydrate composition of tobacco protoplasts transformed with the human UDP-galactose transporter gene hUGT1

　    Horibe T, et al.

样品：植物（烟草）           

  

Genes Brain Behav. 2009 Oct;8(7):650-60.

・    A spontaneous mutation of the Wwox gene and audiogenic seizures in rats with lethal dwarfism and epilepsy.

　    Suzuki H, et al.

样品：大鼠           

 

J Plant Res 2009; 122: 585-95

・    Genetic population structure of Osmunda japonica, rheophilous Osmunda lancea and their hybrids

　    Yatabe Y, et al.

样品：植物           

 

Am J Physiol Renal Physiol 2009; 297: F671-8

・    Npt2a and Npt2c in mice play distinct and synergistic roles in inorganic phosphate metabolism and skeletal development

　    Segawa H, et al.

样品：小鼠           

 

J Immunol 2009;183: 3053-63

・    Crucial contribution of thymic Sirp alpha+ conventional dendritic cells to central tolerance against blood-borne antigens in a CCR2-dependent manner

　    Baba T, et al.

样品：小鼠           

  

J Nat Med 2009; 63: 340-4

・    The botanical origin of kratom (Mitragyna speciosa; Rubiaceae) available as abused drugs in the Japanese markets

　    Maruyama T, et al.

样品：植物         

分析手法：PCR-RFLP法          

 

Emerg Infect Dis 2009;15: 912-5

・    Bartonella quintana in body lice and head lice from homeless persons, San Francisco, California, USA

　    Bonilla DL, et al.

样品：虱子、細菌              

  

Plant Cell Physiol 2009; 50: 1579-86

・    Arabidopsis bile acid:sodium symporter family protein 5 is involved in methionine-derived glucosinolate biosynthesis

　    Sawada Y, et al.

样品：植物（拟南芥）              

 

Mol Genet Metab 2009; 97: 190-5

・    High frequency of acid alpha-glucosidase pseudodeficiency complicates newborn screening for glycogen storage disease type II in the Japanese population

　    Kumamoto S, et al.

样品：滤纸血        

分析手法：ARMS-PCR法         

 

Laboratory Animal research 2009; 25: 75-8

・    Simple Genotyping Method Using Ampdirect Plus and FTA Technologies: Application to the Identification of Transgenic Animals and Their Routine Genetic Monitoring

　    Nakanishi S, et al.

样品：小鼠-大鼠、FTA card              　

 

Int. J. Environ. Res. Public Health 2009; 6: 999-1009

・    Association between a Polymorphism of Aminolevulinate Dehydrogenase (ALAD) Gene and Blood Lead Levels in Japanese Subjects

　    Miyaki K, et al.

样品：人全血              

 

Jpn J Infect Dis 2009; 62: 164-7

・    Direct Colony PCR of Several Medically Important Fungi using Ampdirect(R) Plus

　    Alshahni MM, et al.

样品：真菌（酵母・霉菌）              

 

Biochem. Eng J 2009; 45: 76-81

・    Characterization of bacterial population of raw milk from bovine mastitis by culture-independent

PCR-DGGE method

　    Kuang Y, et al.

样品：牛乳、細菌     

分析手法：PCR-DGGE法          　

 

Lab Chip 2009; 9: 1052-8

・    Microfluidic device using chemiluminescence and a DNA-arrayed thin film transistor photosensor for single nucleotide polymorphism genotyping of PCR amplicons from whole blood

　    Hatakeyama K, et al.

样品：人全血      

分析手法：PCR-RFLP法          

 

Cancer Cell 2009; 15: 195-206

・    Cancer metastasis is accelerated through immunosuppression during Snail-induced EMT of cancer cells

　    Kudo-Saito C, et al.

 

Vet Microbiol 2009; 135: 261-6

・    Detection of cyprinid herpesvirus 3 DNA in river water during and after an outbreak

　    Minamoto T, et al.

样品：河川水、DNA病毒         

 

Methods Mol Biol 2009; 538: 7-27

・    Backtracking of leukemic clones to birth

　    Wiemels J, et al.

样品：新生儿滤纸血           

 

Int J Plant Sci 2009; 170: 237-46

・    Phylogenetic relationship and morphology of Dalzellia gracilis (Podostemaceae, subfamily Tristichoideae)

with proposal of a new genus

　    Koi S, et al.

样品：植物           

 

Infect Dis Clin Pract 2008; 16: 230-4

・    Association of the SLC11A1 Gene Polymorphisms With Susceptibility to Micobacterium Infections in a Japanese Population

　    Asai S, et al.

 

J Vet Diagn Invest 2008; 20: 68-71

・    Molecular screening of canine GM1 gangliosidosis using blood smear specimens after prolonged storage:

detection of carriers among shiba dogs in northern Japan

　    Yamato O, et al.

 

Biosci Biotechnol Biochem 2008; 72: 2831-9

・    Diversity and Similarity of Microbial Communities in Petroleum Crude Oils Produced in Asia

　    Yamane K, et al.

 

Vector Borne Zoonotic Dis 2008; 8: 565-73

・    Detection of Trypanosoma brucei in field-captured tsetse flies and identification of host species fed on by the infected flies

　    Konnai S, et al.

 

J. Pestic Sci 2008; 33: 122-7

・    Environmental distribution and novel high-throughput screening of APEO-degrading bacteria using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-MS)

　    Ichiki Y, et al.

 

Zootaxa 2008; 1746: 15-38

・    Molecular systematics and biogeography of the genus Zizina(Lepidoptera: Lycaenidae)

　    Yago M, et al.

 

Extremophiles 2008; 12: 519-27

・    Phylogenetic and enzymatic diversity of deep subseafloor aerobic microorganisms in organics- and methane-rich sediments off Shimokita Peninsula

　    Kobayashi T, et al.

 

Principles and Technical Aspects of PCR Amplification

・    pp91-101, Springer Netherlands, 2008

　    Elizabeth van Pelt-Verkuil, et al.

 

Zoolog Sci 2008; 25: 838-42

・  Phylogenetic Position of the Endemic Large Carpenter Bee of the Ogasawara Islands, Xylocopa ogasawarensis (Matsumura, 1912) (Hymenoptera: Apidae), Inferred from Four Genes

　    Kawazoe K, et al.

 

Mol Phylogenet Evol 2008; 49: 503-13

・    Redundant species, cryptic host-associated divergence, and secondary shift in Sennertia mites (Acari: Chaetodactylidae) associated with four large carpenter bees (Hymenoptera: Apidae: Xylocopa) in the Japanese island arc

　    Kawazoe K, et al.

 

Odonatologica 2008; 37: 131-44

・    Population genetic differentiaiton in three sympatric damselfly species in a highly fragmented urban landscape

　    Sato M, et al.

 

Blood 2008; 111: 376-8

・    NOTCH1 mutation can be an early, prenatal genetic event in T-ALL

　    Eguchi-Ishimae M, et al.

      

Res Vet Sci 2007; 82: 54-60

・    Nonsense mutation of feline beta-hexosaminidase beta-subunit (HEXB) gene causing Sandhoff disease in a family of Japanese domestic cats

　    Kanae Y, et al.

 

Leuk Res 2007; 31: 1633-40

・    Regulatory polymorphisms of multidrug resistance 1 (MDR1) gene are associated with the development of childhood acute lymphoblastic leukemia

　    Hattori H, et al.

 

Microbiol Immunol 2007; 51: 507-17

・    Plasma levels of unactivated thrombin activatable fibrinolysis inhibitor (TAFI) are down-regulated in young

adult women: analysis of a normal Japanese population

　    Akatsu H, et al.

    

Am J Botany 2007; 94: 1630-41

・    Cryptic species and host specificity in the ectomycorrhizal genus Strobilomyces (Strobilomycetaceae)

　    Sato H, et al.

 

Botanical Journal of the Linnean Society 2007; 155, 1-27

・    A global molecular phylogeny of the fern genus Trichomanes (Hymenophyllaceae) with special reference to stem anatomy

　    Ebihara A, et al.

 

Trop Anim Health Prod 2007; 39: 369-74

・    Comparison of polymerase chain reaction methods for the detection of Theileria equi infection using whole blood compared with pre-extracted DNA samples as PCR templates

　    Alhassan A, et al.

  

Am J Trop Med Hyg 2007; 77: 324-9

・    Establishment of a mass screening method of sand fly vectors for Leishmania infection by molecular biological methods.

　    Kato H, et al

 

Metabolism 2006; 55: 751-7

・    G-protein beta 3 subunit polymorphism C1429T and low-density lipoprotein receptor-related protein 5 polymorphism A1330V are risk factors for hypercholesterolemia in Japanese males–a prospective study over 5 years

　    Suwazono Y, et al.

   

J Vet Med Sci 2006; 68: 27-33.

・    Sequence variation of bovine prion protein gene in Japanese cattle (Holstein and Japanese Black)

　    Nakamitsu S, et al.

  

Ann Hum Genet 2006; 70: 767-77

・    G-protein beta3 subunit variant C825T is a risk factor for hypertension in Japanese females -a prospective cohort study over 5 years

　    Suwazono Y, et al.

  

J Biosci Bioeng 2006; 102: 572-4

・    High-throughput genotyping of filamentous fungus Aspergillus oryzae based on colony direct polymerase chain reaction.

　    Suzuki S, et al

 

Biosci Biotechnol Biochem 2006; 70: 2387-93

・    The molecular phylogeny of the genus Rhizopus based on rDNA sequences.

　    Abe A, et al

 

Appl Environ Microbiol 2006; 72: 7912-5

・    Molecular detection of epiphytic Acaryochloris spp. on marine macroalgae.

　    Ohkubo S, et al

 

Tohoku J Exp Med 2006; 209: 149-57

・    G-protein beta3 subunit gene variant is unlikely to have a significant influence on serum uric acid level in Japanese workers.

　    Suwazono Y, et al

 

Ornithol Sci 2006; 5: 139-143

・    Usefulness of avian buccal cells for molecular sexing.

　    Arima H, et al

 

Int J Mol Med 2006; 17: 77-82

・    The -1438A/G polymorphism in the 5-hydroxytryptamine receptor 2A gene is related to hyperuricemia, increased gamma-glutamyl transpeptidase and decreased high-density lipoprotein cholesterol level in the Japanese population: a prospective cohort study over 5 years.

　    Suwazono Y, et al

 

Arch Virol 2005; 150: 1927-31

・    Rotavirus antigenemia in children with encephalopathy accompanied by rotavirus gastroenteritis

　    Nakagomi T, et al.

    

Thromb Res 2005; 115: 191-7

・    Factor XII Shizuoka, a novel mutation (Ala392Thr) identified and characterized in a patient with congenital coagulation factor XII deficiency

　    Oguchi S, et al.

     

Hemoglobin 2005; 29: 1-10

・     Hb KOCHI [beta141(H19)Leu–>Val (g.1404C–>G); 144-146(HC1-3)Lys-Tyr-His –>0 (g.1413 A–>T)]: a new variant with increased oxygen affinity

　    Miyazaki A, et al.

    

J Epidemiol 2005; 15: 203-10

・     Increased risk of obesity resulting from the interaction between high energy intake and the Trp64Arg polymorphism of the beta3-adrenergic receptor gene in healthy Japanese men.

　    Miyaki K, et al

     

Br J Pharmacol 2005; 145: 818-28

・     Inhibition of the antigen-induced activation of rodent mast cells by putative Janus kinase 3 inhibitors WHI-P131 and WHI-P154 in a Janus kinase 3-independent manner.

　    Watchara Linwong, et al

      

Blood Coagul Fibrinolysis 2004; 15: 367-73

・    Genetic analyses and expression studies identified a novel mutation (W486C) as a molecular basis of congenital coagulation factor XII deficiency

　    Ishii K, et al.

    

Genes Chromosomes Cancer 2004; 39: 335-40

・    Protracted postnatal natural histories in childhood leukemia

　    Maia AT, et al.

   

Obes Res 2004; 12: 4-8

・     Lack of association between human G-protein beta3 subunit variant and overweight in Japanese workers.

　    Suwazono Y, et al

  

Drug Matab. Pharmacokin. 2004; 19: 303-7

・    Genotyping of single nucleotide polymorphism (SNPs) influencing drug response by competitive allele-specific short oligonucleotide hybridization (CASSOH) with immunochromatographic strip.

　    Hiratsuka M, et al

   

J Vet Diagn Invest 2004; 16: 469-72

・    Rapid and simple mutation screening of GM1 gangliosidosis in Shiba dogs by direct amplification of deoxyribonucleic acid from various forms of canine whole-blood specimens.

　    Yamato O, et al

 

      

Blood Coagul Fibrinolysis 2003; 14: 663-70

・    Association of the -159 C –> T polymorphism in the CD14 promoter with variations in serum lipoproteins in healthy subjects.

　    Eilertsen KE, et al.

   

GENES, Chromosomes & Cancer 2003; 37: 36-43

・    Prenatal origin of TEL-AMLI-positive acute lymphoblastic leukemia in children born in California.

　    McHale CM, et al

      

Arch Virol 2002; 147: 2187-95

・    Molecular characterization of serotype G2 and G3 human rotavirus strains that have an apparently identical electropherotype of the short RNA pattern

　    Nakagomi T, et al.

   

BLOOD；2002；99：3801-5

・    In utero origin of t(8;21) AML1-ETO translocations in childhood acute myeloid leukemia

　    Wiemels JL, et al

   

Proc. Natl. Acad. Sci. USA 2002; 99: 15101-6

・    Site-specific translocation and evidence of postnatal origin of the t(1;19) E2A-PBX1 fusion in childhood acute lymphoblastic leukemia.

　    Wiemels JL, et al

  

Clin Lab; 2002; 48: 377-84

・    Various applications of direct PCR using blood samples

　    Nishimura N, et al

     

Proceedings of the Seventh International Colloquium on Paratuberculosis; 2002; 267-9

・    Efficiency of polymerase chain reaction using a novel method of DNA preparation and amplification for detection of Mycobacterium avium subsp. paratuberculosis in bovine fecal samples

　    Kojima K, et al

     

Arch Virol 2001; 146: 557-70

・    Direct evidence for genome segment reassortment between concurrently-circulating human rotavirus strains

　    Watanabe M, et al.

   

Transfusion 2000; 40: 1081-7

・    Elimination of both cell-free and cell-associated HIV infectivity in plasma by a filtration/methylene blue photoinactivation system

　    Abe H, et al.

     

Microbiol Immunol 2000; 44: 957-61

・    Apparent re-emergence of serotype G9 in 1995 among rotaviruses recovered from Japanese children hospitalized with acute gastroenteritis

　    Oka T, et al

   

J Clin Microbiol 2000; 38: 2649-54

・    Major change in the predominant type of “Norwalk-like viruses” in outbreaks of acute nonbacterial gastroenteritis in Osaka city, Japan, between April 1996 and March 1999

　    Iritani N, et al

   

Blood; 2000; 96:264-8

・    Detection of clonotypic IGH and TCR rearrangements in the neonatal blood spots of infants and children with B-cell precursor acute lymphoblastic leukemia

　    Yagi T, et al

     

Ann Clin Biochem; 2000; 37: 674-80

・    Direct PCR from whole blood without DNA isolation

　    Nishimura N, et al

     

Stroke 2000; 31: 2661-4

・    Polymorphism in the promoter of lipopolysaccharide receptor CD14 and ischemic cerebrovascular disease

　    Ito D, et al

  

Stroke 2000; 31: 936-9

・     C242T polymorphism of NADPH oxidase p22 PHOX gene and ischemic cerebrovascular disease in the Japanese population

　    Ito D, et al

  

Stroke 2000; 31: 493-7

・    Association between platelet glycoprotein Iba genotype and ischemic cerebrovascular disease

　    Sonoda A, et al