| 產品名稱 | Acanthamoeba healyi Moura et al. |
|---|---|
| 商品貨號 | B171539 |
| Deposited As | Acanthamoeba culbertsoni Singh and Das |
| Strain Designations | OC-3A [AC-020] |
| Application | characterization of Acanthamoeba polyphaga |
| Biosafety Level | 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 | St Martin's River, MD, 1977 |
| Product Format | freeze-dried |
| Type Strain | no |
| Comments | Taxonomy based on isoenzyme profiles and rDNA PCR-RFLP patterns characterization of Acanthamoeba polyphaga mitochondrial DNA fingerprinting differentiation of Naegleria fowleri from Acanthamoeba using monocolonal antibody phylogeny |
| Medium | ATCC® Medium 712: PYG w/ Additives |
| Growth Conditions | Temperature: 25.0°C Duration: axenic Protocol: ATCCNO: 30010 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium containing 12% (w/v) sucrose to the freeze-dried inner shell vial. Once the culture is completely rehydrated, aseptically add 1 ml of ATCC medium 712 and distribute to a 16 X 125 mm plastic screw-capped test tube or a T-25 tissue culture flask containing 5.0 ml of the same medium. Incubate the test tube culture horizontally with the cap on tight. Trophozoites should be evident in 1-5 days. |
| Subcultivation | Protocol: ATCCNO: 30010 SPEC: This strain is distributed as a freeze-dried preparation. See the general procedures for opening a freeze-dried vial. Aseptically add 0.5 ml of ice cold medium containing 12% (w/v) sucrose to the freeze-dried inner shell vial. Once the culture is completely rehydrated, aseptically add 1 ml of ATCC medium 712 and distribute to a 16 X 125 mm plastic screw-capped test tube or a T-25 tissue culture flask containing 5.0 ml of the same medium. Incubate the test tube culture horizontally with the cap on tight. Trophozoites should be evident in 1-5 days. |
| Cryopreservation | 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. 2.? 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.? 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. 3.? 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.? 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 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation). 6.?? 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 (2-3 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 | TK Sawyer |
| Chain of Custody | ATCC <<--TK Sawyer<<--E.J. Lewis/T.K. Sawyer |
| Year of Origin | 1977 |
| References | Hall J, Voelz H. Bacterial endosymbionts of Acanthamoeba sp.. J. Parasitol. 71: 89-95, 1985. PubMed: 3981353 Gautom RK, et al. Mitochondrial DNA fingerprinting of Acanthamoeba spp. isolated from clinical and environmental sources. J. Clin. Microbiol. 32: 1070-1073, 1994. PubMed: 7913095 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 Flores BM, et al. Differentiation of Naegleria fowleri from Acanthamoeba species by using monoclonal antibodies and flow cytometry. J. Clin. Microbiol. 28: 1999-2005, 1990. PubMed: 2229384 Fritsche TR, et al. Occurrence of bacterial endosymbionts in Acanthamoeba spp. isolated from corneal and environmental specimens and contact lenses. J. Clin. Microbiol. 31: 1122-1126, 1993. PubMed: 8501212 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 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 Alizadeh H, et al. Tear IgA and serum IgG antibodies against Acanthamoeba in patients with Acanthamoeba keratitis. Cornea 20: 622-627, 2001. PubMed: 11473164 Hong YC, et al. Isolation and characterization of a cDNA encoding a mammalian cathepsin L-like cysteine proteinase from Acanthamoeba healyi. Korean J. Parasitol. 40: 17-24, 2002. Flint JA, et al. Genetic analysis of forty isolates of Acanthamoeba Group III by multilocus isoenzyme electrophoresis. Acta Protozool. 42: 317-324, 2003. |
| Cross References | Nucleotide (GenBank) : AF462309 cysteine proteinase (CP1) mRNA, complete coding sequence |
| 梅經理 | 17280875617 | 1438578920 |
| 胡經理 | 13345964880 | 2438244627 |
| 周經理 | 17757487661 | 1296385441 |
| 于經理 | 18067160830 | 2088210172 |
| 沈經理 | 19548299266 | 2662369050 |
| 李經理 | 13626845108 | 972239479 |
