Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University, Wonju, Republic of Korea.
J Clin Microbiol. 2013 May;51(5):1451-7. doi: 10.1128/JCM.01665-12. Epub 2013 Feb 27.
Rapid and accurate identification of the pathogens involved in bloodstream infections is crucial for the prompt initiation of appropriate therapy, as this can decrease morbidity and mortality rates. A PCR-reverse blot hybridization assay for sepsis, the reverse blot hybridization assay (REBA) Sepsis-ID test, was developed; it uses pan-probes to distinguish Gram-positive and -negative bacteria and fungi. In addition, the assay was designed to identify bacteria and fungi using six genus-specific and 13 species-specific probes; it uses additional probes for antibiotic resistance genes, i.e., the mecA gene of methicillin-resistant Staphylococcus aureus (MRSA) and the vanA and vanB genes of vancomycin-resistant enterococci (VRE). The REBA Sepsis-ID test successfully identified clinical isolates and blood culture samples as containing Gram-positive bacteria, Gram-negative bacteria, or fungi. The results matched those obtained with conventional microbiological methods. For the REBA Sepsis-ID test, of the 115 blood culture samples tested, 47 (40.8%) and 49 (42.6%) samples were identified to the species and genus levels, respectively, and the remaining 19 samples (16.5%), which included five Gram-positive rods, were identified as Gram-positive bacteria, Gram-negative bacteria, or fungi. The antibiotic resistances of the MRSA and VRE strains were identified using both conventional microbiological methods and the REBA Sepsis-ID test. In conclusion, the REBA Sepsis-ID test developed for this study is a fast and reliable test for the identification of Gram-positive bacteria, Gram-negative bacteria, fungi, and antibiotic resistance genes (including mecA for MRSA and the vanA and vanB genes for VRE) in bloodstream infections.
快速准确地鉴定血流感染中涉及的病原体对于及时启动适当的治疗至关重要，因为这可以降低发病率和死亡率。开发了一种用于败血症的 PCR-反向斑点杂交分析，即反向斑点杂交分析（REBA）败血症 ID 测试，它使用泛探针来区分革兰氏阳性和革兰氏阴性细菌和真菌。此外，该检测还设计了用于鉴定细菌和真菌的六种属特异性和 13 种种特异性探针；它还使用了针对抗生素耐药基因的额外探针，即耐甲氧西林金黄色葡萄球菌（MRSA）的 mecA 基因和耐万古霉素肠球菌（VRE）的 vanA 和 vanB 基因。REBA 败血症 ID 测试成功地鉴定了临床分离株和血培养样本中含有革兰氏阳性菌、革兰氏阴性菌或真菌。结果与常规微生物学方法一致。对于 REBA 败血症 ID 测试，在测试的 115 个血培养样本中，分别有 47（40.8%）和 49（42.6%）个样本在种和属水平上得到鉴定，其余 19 个样本（16.5%），包括 5 个革兰氏阳性杆菌，被鉴定为革兰氏阳性菌、革兰氏阴性菌或真菌。使用常规微生物学方法和 REBA 败血症 ID 测试鉴定了 MRSA 和 VRE 菌株的抗生素耐药性。总之，本研究开发的 REBA 败血症 ID 测试是一种快速可靠的检测方法，可用于鉴定血流感染中的革兰氏阳性菌、革兰氏阴性菌、真菌和抗生素耐药基因（包括耐甲氧西林金黄色葡萄球菌的 mecA 基因和耐万古霉素肠球菌的 vanA 和 vanB 基因）。
