Abstract
Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, zoonotic infectious diseases caused by Anaplasma phagocytophilum and Ehrlichia chaffeensis, respectively. Early diagnosis is essential for rapid clinical treatment to avoid misdiagnosis and severe patient outcomes. Simple, sensitive and reliable diagnostic methods are urgently needed. In this study, we developed a duplex real-time PCR assay targeting the A. phagocytophilum ankA gene and the E. chaffeensis TRP120 gene, respectively. The lowest limit of detection of the duplex real-time PCR assay was 100 copies of the targeted A. phagocytophilum ankA gene and the E. chaffeensis TRP120 gene per reaction, and the specificity was 100%. Detection in blood DNA samples from the acute stage of illness for 22 HGA cases and 8 HME cases indicated that the duplex real-time PCR assay was more sensitive than the nested PCR assay. The infection of Citellus undulatus Pallas with A. phagocytophilum and E. chaffeensis was first confirmed in Xinjiang Province and the positive rate was 3.1% for A. phagocytophilum, 6.3% for E. chaffeensis and 3.1% for co-infection with both pathogens. The rates of A. phagocytophilum and E. chaffeensis infection of D . silvarum ticks collected from Shanxi Province were 8.2% and 14.8%, respectively, and the co-infection rate was 3.3%. The rates of A. phagocytophilum and E. chaffeensis infection in H. longicornis ticks collected from Shandong Province were 1.6% and 6.3%, respectively, and the co-infection rate was 1.6%.
Highlights
Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, infectious diseases caused by the obligate intracellular bacteria A. phagocytophilum and E. chaffeensis, respectively [1,2,3,4,5]
Among the 64 H. longicornis ticks collected from the Laizhou Bay area, Shandong Province, 1.6% was positive for A. phagocytophilum, and 6.3% were positive for E. chaffeensis; and the co-infection rate was 1.6% (Table 4)
E. chaffeensis TRP120 proteins are strongly recognized by the host immune response, and recombinant TRP120 protein is considered to be a valuable diagnostic antigen
Summary
Human granulocytic anaplasmosis (HGA) and human monocytic ehrlichiosis (HME) are emerging, tick-borne, infectious diseases caused by the obligate intracellular bacteria A. phagocytophilum and E. chaffeensis, respectively [1,2,3,4,5]. As an emerging tick-borne infectious disease, a survey of farm workers in rural areas of Tianjin has revealed that the average seroprevalence of anaplasmosis among the population is 8.8% [16]. Misdiagnoses may result in severe multiorgan symptoms or even death in healthy adults and children, despite the availability of low-cost, effective antimicrobial therapy In this regard, early diagnosis is essential for rapid clinical treatment [19]. Co-infection by E. chaffeensis and A. phagocytophilum has been frequently reported in both clinical patients and ticks in recent years in China [20]. This method could be widely applied to test various samples such as patient and domestic animal blood and tissue samples as well as tick samples, and the specificity, sensitivity and reproducibility of the real-time PCR assay are not influenced by background DNA
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