![A-1 [AC-001]](/static/images/no_picture.jpg)
Acanthamoeba culbertsoni Singh and Das 拉丁名
(ATCC® 30171™) 統(tǒng)一編號(hào)
Strain Designations 別名 A-1 [AC-001]
Application 用途 1,3 diamnopropane production ；
Characterization of Acanthamoeba polyphaga ；
PCR for identification of Naegleria fowleri
Biosafety Level 生物安全等級(jí) 2
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. Isolation Primary monkey kidney tissue culture, India (?), 1957
Product Format 提供形式 frozen
Storage Conditions 生長條件
Frozen Cultures: -70°C for 1 week; liquid N2 vapor for long term storage
Freeze-dried Cultures: 2-8°C
Live Cultures: See Protocols section for handling information
Type Strain no
Antibiotic Resistance resistant to complement lysis
Comments Taxonomy based on isoenzyme profiles and rDNA PCR-RFLP patterns； Two genetic markers that distinguish pathogenic and nonpathogenic strains； Resistant to complement lysis； Species description； Antiamoebic action of diamidines； 1,3 diamnopropane production； Specific antigens revealed by protein immunoblotting； Characterization of Acanthamoeba polyphaga； Scanning electron microscopy； PCR for identification of Naegleria fowleri； Review； Phylogeny；
Medium ATCC® Medium 712: PYG w/ Additives
Growth Conditions 生長條件
Temperature: 25°C
Culture System: Axenic
Cryopreservation Harvest and Preservation
1，To achieve the best results set up cultures with several different inocula (e.g. 0.25 mL, 0.5 mL, 1.0 mL). Harvest cultures and pool when the culture that received the lowest inoculum is at or near peak density. If the cell concentration exceeds the required level do not centrifuge, but adjust the concentration to between 2 x 106 and 2 x 107cysts/mL with fresh medium.
2，If the concentration is too low, centrifuge at 600 x g for 5 min and resuspend the pellet in the volume of fresh medium required to yield the desired concentration. While cells are centrifuging prepare a 15% (v/v) solution of sterile DMSO as follows: Add the required volume of DMSO to a glass screw-capped test tube and place it in an ice bath. Allow the DMSO to solidify.
3，Add the required volume of refrigerated medium. Dissolve the DMSO by inverting the tube several times. Note: If the DMSO solution is not prepared on ice, an exothermic reaction will occur that may precipitate certain components of the medium.
4，Mix the cell preparation and the DMSO in equal portions. Thus, the final concentration will be between 106 and 107 cells/mL and 7.5% (v/v) DMSO. The time from the mixing of the cell preparation and DMSO stock solution before the freezing process is begun should be no less than 15 min and no longer than 30 min.
5，Dispense in 0.5 mL aliquots into 1.0 mL to 2.0 mL sterile plastic screw-capped cryules (special plastic vials for cryopreservation). Place the vials in a controlled rate freezing unit.
From room temperature cool at -1°C/min to -40°C.
If the freezing unit can compensate for the heat of fusion, maintain rate at -1°C/min through the heat of fusion. At -40°C plunge into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus. Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen. (The cooling rate in this apparatus is approximately -1°C/min.)
7，The frozen preparations are stored in either the vapor or liquid phase of a nitrogen freezer.
8，To establish a culture from the frozen state place an ampule in a water bath set at 35°C (2 to 3 min). Immerse the vial just sufficient to cover the frozen material. Do not agitate the vial.
9，Immediately after thawing, aseptically remove the contents of the ampule and inoculate into 5 mL of fresh ATCC medium 712 in a T-25 tissue culture flask or plastic 16 x 125 mm screw-capped test tube. Incubate at 25°C.
Name of Depositor CG Culbertson
Year of Origin 1957
References 參考文獻(xiàn) Allen DJ, Didio LJ. Acanthamoeba culbertsoni as revealed in scanning electron microscopy. Ohio J. Sci. 76: 167-171, 1976. Am. J. Clin. Pathol. 35: 195-202, 1961.
Daggett PM, et al. Distribution and possible interrelationships of pathogenic and nonpathogenic Acanthamoeba from aquatic environments. Microb. Ecol. 8: 371-386, 1982.
Daggett PM, et al. A molecular approach to the phylogeny of Acanthamoeba. Biosystems 18: 399-405, 1985. PubMed: 4084681
Flint JA, et al. Genetic analysis of forty isolates of Acanthamoeba Group III by multilocus isoenzyme electrophoresis. Acta Protozool. 42: 317-324, 2003. Gast RJ. Development of an Acanthamoeba-specific reverse dot-blot and the discovery of a new ribotype. J. Eukaryot. Microbiol. 48: 609-615, 2001. PubMed: 11831768
Hall J, Voelz H. Bacterial endosymbionts of Acanthamoeba sp.. J. Parasitol. 71: 89-95, 1985. PubMed: 3981353
Howe D, et al. Identification of two genetic markers that distinguish pathogenic and nonpathogenic strains of Acanthamoeba spp.. Parasitol. Res. 83: 435-348, 1997. PubMed: 9197389
John DTOpportunistically pathogenic free-living amebaeIn: John DTParasitic protozoa2nd ed.3San DiegoAcademic Presspp. 143-246, 1993 Kilvington S, Beeching J. Development of a PCR for identification of Naegleria fowleri from the environment. Appl. Environ. Microbiol. 61: 3764-3767, 1995. PubMed: 7487014
Kim YH, et al. Close relatedness of Acanthamoeba pustulosa with Acanthamoeba palestinensis based on isoenzyme profiles and rDNA PCR-RFLP patterns. Korean J. Parasitol. 34: 259-266, 1996. PubMed: 9017912
Kishore P, Shukla OP. Med. Sci. Res. 17: 601-602, 1989.
Kong HH, et al. Mitochondrial DNA restriction fragment length polymorphism (RFLP) and 18S small-subunit ribosomal DNA PCR-RFLP analyses of Acanthamoeba isolated from contact lens storage cases of residents in southwestern Korea. J. Clin. Microbiol. 40: 1199-1206, 2002. PubMed: 11923331
Marciano-Cabral F, Toney DM. The interacion of Acanthamoeba spp. with activated macrophages and with macrophage cell lines. J. Eukaryot. Microbiol. 45: 452-458, 1998. PubMed: 9703682
Moura H, et al. Acanthamoeba healyi n. sp. and the isoenzyme and immunoblot profiles of Acanthamoeba spp., groups 1 and 3. J. Protozool. 39: 573-583, 1992. PubMed: 1522539 Marciano-Cabral F, Cabral G. Acanthamoeba spp. as agents of disease in humans. Clin. Microbiol. Rev. 16: 273-307, 2003. PubMed: 12692099 Pettit DA, et al. In vitro destruction of nerve cell cultures by Acanthamoeba spp.: a transmission and scanning electron microscopy study. J. Parasitol. 82: 769-777, 1996. PubMed: 8885887
Powell EL, et al. Identification of antigens of pathogenic free-living amoebae by protein immunoblotting with rabbit immune and human sera. Clin. Diagn. Lab. Immunol. 1: 493-499, 1994. PubMed: 8556491
Reveiller FL, et al. Isolation of a unique membrane protein from Naegleria fowleri. J. Eukaryot. Microbiol. 48: 676-682, 2001. PubMed: 11831777
Sawyer TK, et al. Acanthamoeba jacobsi sp. n. (Protozoa: Acanthamoebidae) from sewage contaminated ocean sediments. J. Helminthol. Soc. Wash. 59: 223-226, 1992.
Shukla OP, et al. Identification of the polyamine N8-acetyltransferase involved in the pathway of 1,3-diaminopropane production in Acanthamoeba culbertsoni. Parasitol. Res. 82: 270-272, 1996. PubMed: 8801564
Stothard DR, et al. The evolutionary history of the genus Acanthamoeba and the identification of eight new 18S rRNA gene sequence types. J. Eukaryot. Microbiol. 45: 45-54, 1998. PubMed: 9495032
Stothard DR, et al. Fluorescent oligonucleotide probes for clinical and environmental detection of Acanthamoeba and the T4 18S rRNA gene sequence type. J. Clin. Microbiol. 37: 2687-2693, 1999. PubMed: 10405422
Toney DM, Marciano-Cabral F. Resistance of Acanthamoeba species to complement lysis. J. Parasitol. 84: 338-344, 1998. PubMed: 9576508
Cross References Nucleotide (GenBank) : AF019067 Acanthamoeba culbertsoni Lilly A-1 18S ribosomal RNA gene, partial sequence.
ATCC30171
