Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Infectious Diseases Unit, Internal Medicine Department, Universiti Teknologi MARA, Sungai Buloh, Selangor, Malaysia.
PLoS One. 2020 Sep 11;15(9):e0239069. doi: 10.1371/journal.pone.0239069. eCollection 2020.
The diagnosis of leptospirosis remains a challenge due to its non-specific symptoms and the biphasic nature of the illness. A comprehensive diagnosis that includes both molecular (polymerase chain reaction (PCR)) and serology is vital for early detection of leptospirosis and to avoid misdiagnosis. However, not all samples could be subjected to both tests (serology and molecular) due to budget limitation, infrastructure, and technical expertise at least in resource-limited countries. We evaluated the usefulness of testing the clinically suspected leptospirosis cases with both techniques on all samples collected from the patients on the day of admission. Among the 165 patient's blood/serum samples tested (from three hospitals in Central Malaysia), 43 (26%) showed positivity by microscopic agglutination test (MAT), 63 (38%) by PCR, while 14 (8%) were positive by both MAT and PCR. For PCR, we tested two molecular targets (lipL32 by qPCR and 16S rDNA or rrs by nested PCR) and detected lipL32 in 47 (29%) and rrs gene in 63 (38%) patients. The use of more than one target gene for PCR increased the detection rates. Hence, a highly sensitive multiplex PCR targeting more than one diagnostic marker is recommended for the early detection of Leptospira in suspected patients. When the frequencies for positivity detected either by MAT or PCR combined, leptospirosis was diagnosed in a total of 92 (56%) patients, a higher frequency compared to when samples were only tested by a single method (MAT or PCR). The results from this study suggest the inclusion of both serology and molecular methods for every first sample irrespective of the days post-onset of symptoms (DPO) collected from patients for early diagnosis of leptospirosis.
由于其非特异性症状和疾病的双相性质，钩端螺旋体病的诊断仍然具有挑战性。全面的诊断包括分子（聚合酶链反应（PCR））和血清学对于早期发现钩端螺旋体病和避免误诊至关重要。然而，由于预算限制、基础设施和技术专业知识，并非所有样本都可以同时进行这两种测试（血清学和分子学），至少在资源有限的国家是这样。我们评估了在入院当天对所有患者样本同时使用这两种技术检测临床疑似钩端螺旋体病病例的有用性。在对来自马来西亚中部 3 家医院的 165 名患者的血液/血清样本进行检测中，43 份（26%）显微镜凝集试验（MAT）阳性，63 份（38%）PCR 阳性，14 份（8%）MAT 和 PCR 均阳性。对于 PCR，我们测试了两个分子靶标（qPCR 的 lipL32 和嵌套 PCR 的 16S rDNA 或 rrs），并在 47 名（29%）和 63 名（38%）患者中检测到 lipL32 和 rrs 基因。使用多个靶基因进行 PCR 可提高检测率。因此，建议使用针对多个诊断标志物的高灵敏度多重 PCR 来早期检测疑似患者中的钩端螺旋体。当 MAT 或 PCR 联合检测的阳性率相加时，总共诊断出 92 名（56%）患者患有钩端螺旋体病，这比仅使用单一方法（MAT 或 PCR）检测时的频率更高。本研究结果表明，无论从患者采集的症状出现后第几天（DPO），都应将血清学和分子学方法纳入每个初次样本，以早期诊断钩端螺旋体病。
