Department of Biotechnology, College of Life Science and Biotechnology, Yonsei University, 50 Yonsei-ro, Seodaemun-gu, Seoul 03722, Republic of Korea.
Division of Infectious Diseases, Department of Internal Medicine, Pusan National University Yangsan Hospital, Yangsan 50561, Republic of Korea.
Biosensors (Basel). 2024 Oct 10;14(10):493. doi: 10.3390/bios14100493.
Scrub typhus is caused by the Gram-negative obligate intracellular bacterium , and this tick-borne disease is difficult to distinguish from other acute febrile illnesses as it typically presents with symptoms such as rash, crusting at the bite site, headache, myalgia, lymphadenopathy, and elevated liver transaminases. It can often be diagnosed clinically, but not all patients present with characteristic symptoms, so serological diagnosis and molecular techniques may be required. However, existing diagnostic tests often have low sensitivity and specificity, making early detection difficult. This study presents a nucleic acid extraction method using large volumes of plasma and buffy coat to increase sensitivity, as well as an improved detection method using two target genes. Using the I-PULL device, nucleic acids can be extracted from up to 4 mL of sample in 30 min, avoiding contamination. The extracted DNA detects two genes of , increasing sensitivity compared to single-gene detection. Clinical validation in 38 patient samples showed 100% specificity and 95.24% sensitivity for the single target gene, with specificity and sensitivity rising to 100% when both genes are analyzed. This molecular diagnostic platform can be useful for distinguishing scrub typhus from similar diseases.
恙虫病是由革兰氏阴性专性细胞内细菌引起的，这种由蜱传播的疾病很难与其他急性发热疾病区分，因为它通常表现为皮疹、叮咬部位结痂、头痛、肌痛、淋巴结病和肝转氨酶升高。它通常可以通过临床诊断，但并非所有患者都有特征性症状，因此可能需要血清学诊断和分子技术。然而，现有的诊断测试通常灵敏度和特异性较低，使得早期检测变得困难。本研究提出了一种使用大量血浆和白细胞层进行核酸提取的方法，以提高灵敏度，并提出了一种使用两个靶基因的改进检测方法。使用 I-PULL 装置，可在 30 分钟内从多达 4 毫升的样本中提取核酸，避免污染。提取的 DNA 可检测到两个基因，与单基因检测相比，灵敏度提高。对 38 个患者样本的临床验证显示，单靶基因的特异性和灵敏度均为 100%，当分析两个基因时，特异性和灵敏度均提高至 100%。这种分子诊断平台可用于区分恙虫病和类似疾病。
