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CHROMagar Candida Medium for Direct Susceptibility Testing of Yeast from Blood Cultures

机译:CHROMagar念珠菌培养基可直接用于血液培养酵母的药敏试验

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摘要

An evaluation was performed on 95 blood cultures positive for Candida spp. to determine the correlation of direct susceptibility testing of fluconazole versus both standardized disk diffusion and MIC methods. For direct testing, an aliquot taken from BD BACTEC Plus and/or BD BACTEC Lytic/10 bottles (Becton Dickinson [BD], Sparks, MD) positive by gram stain for yeast was subcultured to CHROMagar Candida (BD), and a 25-μg fluconazole disk (BD) was placed on the plate. The area of growth inhibition surrounding the disk was measured at 24 and 48 h. In addition, a subculture of the isolate was tested by a microdilution MIC using YeastOne (TREK Diagnostics Systems Inc., OH) and disk diffusion (NCCLS M44-A) using a standardized inoculum plated onto CHROMagar Candida as well as Mueller-Hinton agar to which 2% glucose and 0.5 μg/ml methylene blue dye was added (MH-GMB). The categorical interpretation derived from the MIC was used as the reference to which the disk diffusion results were compared. There were a total of 41 Candida albicans, 23 Candida glabrata, 20 Candida parapsilosis, 9 Candida tropicalis, and 1 each of Candida krusei and Candida lusitaniae tested. At 24 h there was full agreement among the methods for all C. albicans, C. tropicalis, C. lusitaniae, and C. krusei isolates. For the C. parapsilosis isolates at 24 h there was one very major discrepancy using the direct CHROMagar and one major error with the standardized MH-GMB. The majority of the errors were seen at 24 h with the C. glabrata isolates. Of the 23 C. glabrata isolates at 24 h by direct CHROMagar, there were 10 minor and 1 very major error; by MH-GMB there were 12 minor and 2 very major errors; and by standardized CHROMagar Candida there were 13 minor and 2 major errors. There were no very major errors with C. glabrata when all plates were read at 48 h. At 24 h by the direct and standardized CHROMagar the majority of C. glabrata isolates were more resistant, whereas by MH-GMB they were more susceptible than the reference MIC interpretation. In summary, subculturing yeast directly from blood cultures onto CHROMagar to which a fluconazole disk has been added may provide a presumptive identification at 24 h and, with the exception of C. glabrata, was able to predict the susceptibility to fluconazole with the majority of Candida isolates examined in this evaluation.
机译:对假丝酵母念珠菌阳性的95种血液培养物进行了评估。以确定氟康唑的直接药敏试验与标准化磁盘扩散法和MIC方法的相关性。为了进行直接测试,将对酵母呈革兰氏染色呈阳性的BD BACTEC Plus和/或BD BACTEC Lytic / 10瓶(Becton Dickinson [BD],Sparks,MD)中提取的等分试样传代至CHROMagar Candida(BD),然后将25-将μg氟康唑片(BD)放在板上。在24和48小时测量围绕盘的生长抑制面积。此外,使用YeastOne(TREK Diagnostics Systems Inc.,OH)进行微量稀释MIC,并使用接种在CHROMagar Candida和Mueller-Hinton琼脂上的标准接种物,通过圆盘扩散法(NCCLS M44-A)对分离株的传代培养进行测试。加入2%葡萄糖和0.5μg/ ml亚甲基蓝染料(MH-GMB)。来自MIC的分类解释被用作参考,比较了磁盘扩散结果。总共测试了41个白色念珠菌,23个光滑念珠菌,20个副念珠菌,9个热带念珠菌以及krusei和lusitaniae念珠菌各1个。在24小时时,所有白色念珠菌,热带念珠菌,卢梭菌和克鲁斯梭菌的分离方法完全一致。对于在24 h时副枝梭菌的分离株,使用直接的CHROMagar存在一个非常重大的差异,而使用标准的MH-GMB则存在一个重大错误。大多数错误是在glabrata C. glabrata分离株于24 h见到的。直接CHROMagar在24 h分离出的23株毛孢梭菌中,有10个轻微错误和1个非常重大错误; MH-GMB报告有12个小错误和2个非常大的错误;通过标准化的CHROMagar Candida,有13个小错误和2个大错误。在48 h读取所有平板时,没有很大的错误。在直接和标准化的CHROMagar的24小时内,大多数的光滑念珠菌分离株具有更高的抗药性,而通过MH-GMB,它们比参考MIC解释更易感。总而言之,将酵母直接从血液培养物传代到已添加氟康唑盘的CHROMagar上,可以在24 h时提供推测鉴定,除了 C。 glabrata 能够预测此次评估中检测到的大多数 Candida 分离株对氟康唑的敏感性。

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