斑疹伤寒(ST)是一种被忽视的急性,发热,由胞内寄生虫东方虫引起的传染病,立克次体科的革兰氏阴性球杆菌。早期和精确的诊断对于降低发生疾病并发症的风险至关重要。然而,IgM抗体酶联免疫吸附测定(IgMELISA)和间接免疫荧光测定(IFA)仍然是诊断的关键。然而,由于在诊断准确性研究中需要统一的方法来确定不同地理位置的适当ELISA截止值,因此这可能对早期诊断更有帮助。因此,我们的目的是使用巢式PCR(nPCR)和DNA序列分析作为早期诊断的分子标记来研究O.tu虫类型特异性56千道尔顿(kDa)蛋白基因。在10439例疑似病例中,1147/10439(11%)例患者IgMELISA阳性。nPCR后随机检测了1044/10,439(10%)个样品,并与IgMELISA结果和DNA序列分析进行了比较。使用巢式PCR和IgMELISA方法,13%(134/1044)和12%(125/1044)的样本呈阳性,分别。血清学方法无法复制nPCR证明的大量阳性病例;因此,这两种技术的显著互斥需要进一步研究。此外,我们的系统发育分析揭示了具有Karp相关菌株的分离株聚集,提供对传输动力学的洞察。因此,分子诊断方法可能有助于感染的早期诊断,并能够在流行地区迅速治疗ST。我们的结果表明,IgMELISA可以与nPCR结合提供完整的诊断优势,并且可以成为准确诊断的重要工具。此外,样品的DNA测序分析表明,Karp相关菌株是主要菌株。此外,需要对来自各个地区的样本进行研究,并结合对O.tsugamushi进行全基因组测序,以更好地了解感染机制并开发强大的早期检测方法。
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Scrub typhus (ST) is a neglected acute, febrile, infectious disease caused by the intracellular parasite Orientia tsutsugamushi, a gram-negative coccobacillus of the family Rickettsiaceae. Early and precise diagnosis is crucial to reduce the risk of developing disease complications. However, IgM antibody enzyme-linked immunosorbent assay (IgM ELISA) and indirect immunofluorescence assay (IFA) remain essential for diagnosis. However, it could be more helpful for early diagnosis due to the need for uniformity of approach in the diagnostic accuracy studies to determine appropriate ELISA cut-offs for various geographic locations. Hence, we aim to
study the O. tsutsugamushi type-specific 56 kilodalton (kDa) protein gene using nested PCR (nPCR) and DNA sequence analysis as a molecular marker for early diagnosis. Out of 10,439 suspected cases, 1147/10,439 (11%) patients were positive for IgM ELISA. A total of 1044/10,439 (10%) samples were randomly tested after nPCR and compared with IgM ELISA results and DNA sequence analysis. Using nested PCR and IgM ELISA methods, 13% (134/1044) and 12% (125/1044) of the samples were positive, respectively. The serology method could not replicate the substantial number of positive cases demonstrated by nPCR; therefore, significant mutual exclusivity of the two techniques requires further investigation. Furthermore, our phylogenetic analysis revealed a clustering of isolates with Karp-related strains, providing insight into the transmission dynamics. Therefore, molecular diagnostic methods may aid in the early diagnosis of infection and enable prompt treatment of ST in endemic regions. Our results show that IgM ELISA can provide complete diagnostic advantages in conjunction with nPCR and can be an essential tool for accurate diagnosis. In addition, the DNA sequencing analysis of the samples showed that Karp-related strains were the main strains. Furthermore, research with samples from various regions in combination with the entire genome sequencing of O. tsutsugamushi is required to understand the infection mechanism better and develop robust early detection methods.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s00580-023-03443-8.