Graczyk T K, Fayer R, Cranfield M R, Owens R
Department of Molecular Microbiology and Immunology, School of Hygiene and Public Health, Johns Hopkins University, Baltimore, Maryland 21205, USA.
J Parasitol. 1997 Feb;83(1):111-4.
Cryptosporidium parvum oocysts infectious to neonatal BALB/c mice were processed by the cellulose-acetate membrane (CAM) filter dissolution method to determine if the procedure that utilizes acetone incubation and alcohol centrifugations alters their viability (determined by in vitro excystation) or infectivity (determined by infectivity bioassay). In addition, most oocysts with altered viability by desiccation, heat inactivation, and snap freezing that were processed by the CAM filter dissolution method were nonrefractile, unstained oocyst ghosts. The remaining organisms, oocyst shells, were lightly stained with the acid-fast stain. Infectious oocysts retained their infectivity and nonviable oocysts (oocyst shells) retained their morphology when processed by the CAM dissolution method. Infectious oocysts, oocyst shells, and oocyst ghosts produced positive reactions of similar intensity in direct immunofluorescence antibody staining, utilizing the MERIFLUOR Cryptosporidium/Giardia test kit. Cryptosporidium oocysts recovered from finished drinking water by the CAM dissolution method can be subjected to testing for their viability and infectivity.
采用醋酸纤维素膜(CAM)滤膜溶解法处理对新生BALB/c小鼠具有感染性的微小隐孢子虫卵囊,以确定利用丙酮孵育和酒精离心的程序是否会改变其活力(通过体外脱囊确定)或感染性(通过感染性生物测定确定)。此外,大多数经CAM滤膜溶解法处理的因干燥、热灭活和速冻而活力改变的卵囊是非折光性、未染色的卵囊空壳。其余的生物体,即卵囊壳,经抗酸染色后轻度染色。经CAM溶解法处理时,感染性卵囊保持其感染性,无活力的卵囊(卵囊壳)保持其形态。利用MERIFLUOR隐孢子虫/贾第虫检测试剂盒,感染性卵囊、卵囊壳和卵囊空壳在直接免疫荧光抗体染色中产生强度相似的阳性反应。通过CAM溶解法从成品饮用水中回收的隐孢子虫卵囊可进行活力和感染性检测。