Varettas Kerry, Mukerjee Chinmoy, Taylor Peter C
Division of Microbiology, SEALS, St. George Hospital, Kogarah, New South Wales, Australia.
J Clin Microbiol. 2005 Sep;43(9):4613-5. doi: 10.1128/JCM.43.9.4613-4615.2005.
The tube coagulase test (TCT) performed directly from positive blood culture bottles has been used to reduce the turnaround time for identifying Staphylococcus aureus. Most reports have shown the test to be specific but often lacking sufficient sensitivity to be useful. In a prospective study of blood culture bottles (BCB) signaling positive, with a Gram-stained smear showing gram-positive cocci resembling staphylococci, the sensitivity of the direct TCT was improved by diluting the BCB broth 1:10 in saline before inoculating 0.1 ml into 1.0 ml of 10% pooled human plasma. It was hypothesized that the improved sensitivity might be explained by reduced carryover of the anticoagulant sodium polyanetholesulfonate (SPS) used in blood culture media. By titrating the inoculum size and the concentration of SPS in an in vitro checkerboard assay, it was shown that concentrations of SPS >0.0008% prevented plasma coagulation. The 1:10 dilution of blood culture broth reduced the amount of residual SPS carried over to the TCT to a level (0.0005%) that did not impair plasma coagulation. The direct TCT inoculated with a 1:10 saline dilution of blood culture broth achieved 100% specificity and sensitivity within 4 h of inoculation without reducing the quality or quantity of coagulum.
直接从阳性血培养瓶进行的试管凝固酶试验(TCT)已被用于缩短鉴定金黄色葡萄球菌的周转时间。大多数报告显示该试验具有特异性,但往往缺乏足够的敏感性以发挥作用。在一项对血培养瓶(BCB)呈阳性信号且革兰氏染色涂片显示革兰氏阳性球菌类似葡萄球菌的前瞻性研究中,通过将BCB肉汤用生理盐水按1:10稀释,然后将0.1 ml接种到1.0 ml 10%混合人血浆中,直接TCT的敏感性得到了提高。据推测,敏感性提高可能是由于血培养培养基中使用的抗凝剂聚茴香脑磺酸钠(SPS)的残留减少所致。通过在体外棋盘试验中滴定接种量和SPS浓度,结果表明SPS浓度>0.0008%可阻止血浆凝固。血培养肉汤1:10稀释可将带入TCT的残留SPS量降低到不影响血浆凝固的水平(0.0005%)。接种血培养肉汤1:10生理盐水稀释液的直接TCT在接种后4小时内实现了100%的特异性和敏感性,且不降低凝块的质量或数量。