Hamilton J R, Noble A, Denning D W, Stevens D A
Department of Pathology, Santa Clara Valley Medical Center, San Jose, California.
J Clin Microbiol. 1991 Feb;29(2):333-9. doi: 10.1128/jcm.29.2.333-339.1991.
Cryptococcal antigen titers in 97 serum and 42 cerebrospinal fluid (CSF) specimens from 37 AIDS patients with culture-proven cryptococcal infection were determined with the Meridian kit (Meridian Diagnostics Inc., Cincinnati, Ohio) before and after treatment with pronase. The geometric mean titers before and after pronase treatment were 1:45 and 1:588 in serum and 1:97 and 1:79 in CSF, respectively. Only on serum (but not CSF) specimens after pronase treatment were (i) titers increased by 2 to 13 dilutions on 57% of the specimens, all of which had titers of less than or equal to 1:128 before pronase treatment, (ii) false-negative reactions on 27% of specimens before pronase treatment eliminated, all of which had titers from 1:4 to 1:4,096, (iii) prozone-like reactions (titer, less than or equal to 1:256) on 9% of the specimens before pronase treatment eliminated, and (iv) agglutination reactions on all specimens stronger and easier to interpret. Antifungal agents added to serum as well as freeze-thaw cycles did not change antigen titers in serum. After two separate tests, the same titers were obtained on 94% of 35 serum specimens that were treated with pronase and on 96% of 53 CSF specimens that were not treated with pronase. A total of 26 serum specimens and 28 CSF specimens from patients with no cryptococcal disease were negative before and after pronase treatment. The IBL kit (International Biological Labs Inc., Cranbury, N.J.) was compared with the Meridian kit on 41 serum specimens and 14 CSF specimens. Results from the two kits agreed on 54 and 68% of serum specimens and 86 and 93% of CSF specimens before and after pronase treatment, respectively. The IBL kit generally produced higher titers on specimens in disagreement and produced no prozone-like reactions. Routine pronase treatment of serum is recommended with the Meridian kit in order to eliminate false-negative and unclear agglutination reactions by producing a consistent interpretation of agglutination reactions. CSF specimens do not require pronase treatment. Titer results produced by the kits from the two different manufacturers varied considerably: the kits should not be used interchangeably for determining antigen titers in serum specimens.
使用Meridian试剂盒(Meridian诊断公司,俄亥俄州辛辛那提)测定了37例经培养证实患有隐球菌感染的艾滋病患者的97份血清和42份脑脊液(CSF)标本在链霉蛋白酶处理前后的隐球菌抗原滴度。链霉蛋白酶处理前后血清中的几何平均滴度分别为1:45和1:588,脑脊液中的几何平均滴度分别为1:97和1:79。仅在链霉蛋白酶处理后的血清(而非脑脊液)标本中,(i)57%的标本滴度增加了2至13个稀释度,所有这些标本在链霉蛋白酶处理前的滴度均小于或等于1:128;(ii)消除了链霉蛋白酶处理前27%标本的假阴性反应,所有这些标本的滴度为1:4至1:4096;(iii)消除了链霉蛋白酶处理前9%标本的前带样反应(滴度小于或等于1:256);(iv)所有标本的凝集反应更强且更易于判读。添加到血清中的抗真菌剂以及冻融循环均未改变血清中的抗原滴度。经过两次单独测试,在35份经链霉蛋白酶处理的血清标本中有94%以及在53份未经链霉蛋白酶处理的脑脊液标本中有96%获得了相同的滴度。来自无隐球菌病患者的26份血清标本和28份脑脊液标本在链霉蛋白酶处理前后均为阴性。在41份血清标本和14份脑脊液标本上对IBL试剂盒(国际生物实验室公司,新泽西州克兰伯里)与Meridian试剂盒进行了比较。在链霉蛋白酶处理前后,两种试剂盒在血清标本中的结果一致性分别为54%和68%,在脑脊液标本中的结果一致性分别为86%和93%。对于结果不一致的标本,IBL试剂盒通常产生更高的滴度,且未产生前带样反应。建议使用Meridian试剂盒对血清进行常规链霉蛋白酶处理,以便通过对凝集反应产生一致的判读来消除假阴性和不明确的凝集反应。脑脊液标本不需要链霉蛋白酶处理。来自两个不同制造商的试剂盒产生的滴度结果差异很大:在测定血清标本中的抗原滴度时,两种试剂盒不可互换使用。