Hodes D S, Heon D, Hass A, Hyatt A C, Hodes H L
Department of Pediatrics, Mount Sinai School of Medicine, City University of New York, New York 10029.
J Clin Microbiol. 1987 Sep;25(9):1701-4. doi: 10.1128/jcm.25.9.1701-1704.1987.
A commercially available endotoxin assay (CS-TAL) employing a chromogenic peptide and an amebocyte lysate from the Japanese horseshoe crab, Tachypleus tridentatus, gave a positive result with aqueous extracts of all 15 strains of Candida albicans and 1 strain each of Candida tropicalis, Cryptococcus neoformans, and a Mucor species that we tested. Purified glucans prepared from the Candida strains gave the same results. Reconstruction experiments showed that the positive results were not due to contaminating endotoxin. By contrast, assays employing amebocyte lysates of the American horseshoe crab, Limulus polyphemus, were inconsistent. Japanese workers have presented evidence that glucans activate the Tachypleus amebocyte lysate system by acting on an enzyme different from that on which endotoxin acts. Using a Tachypleus lysate preparation (Endospecy; Seikagaku Kogyo, Tokyo, Japan) from which this enzyme was excluded, we demonstrated a 5- to 10-fold drop in reactivity to the aqueous Candida extracts and glucans, whereas reactivity to endotoxin was unchanged. Normal human plasma was shown to decrease the effect of fungal extracts on CS-TAL. This inhibition was completely removed by heating the plasma. Our results suggest that Tachypleus systems may be of use clinically in distinguishing bacterial from fungal infections.
一种市售的内毒素检测方法(CS-TAL),采用一种显色肽和来自日本鲎(Tachypleus tridentatus)的变形细胞溶解物,对我们检测的所有15株白色念珠菌以及热带念珠菌、新型隐球菌和一种毛霉属菌株各1株的水提取物均给出了阳性结果。从念珠菌菌株制备的纯化葡聚糖也得到了相同的结果。重建实验表明,阳性结果并非由于内毒素污染。相比之下,采用美洲鲎(Limulus polyphemus)变形细胞溶解物的检测方法结果并不一致。日本研究人员已提出证据表明,葡聚糖通过作用于一种不同于内毒素作用的酶来激活鲎变形细胞溶解物系统。使用一种排除了该酶的鲎溶解物制剂(Endospecy;日本东京生化学工业株式会社),我们证明对念珠菌水提取物和葡聚糖的反应性下降了5至10倍,而对内毒素的反应性未变。结果显示,正常人血浆可降低真菌提取物对CS-TAL的影响。通过加热血浆可完全消除这种抑制作用。我们的结果表明,鲎检测系统可能在临床上有助于区分细菌感染和真菌感染。