Gegembauer Gregory, Araujo Leticia Mendes, Pereira Edy Firmina, Rodrigues Anderson Messias, Paniago Anamaria Mello Miranda, Hahn Rosane Christine, de Camargo Zoilo Pires
Universidade Federal de São Paulo (UNIFESP), Departamento de Microbiologia, Imunologia e Parasitologia, Disciplina de Biologia Celular, São Paulo, São Paulo, Brasil.
Faculdade de Medicina (FAMED) da Universidade Federal de Mato Grosso do Sul (UFMS), Campo Grande, Mato Grosso do Sul, Brasil.
PLoS Negl Trop Dis. 2014 Jul 17;8(7):e2986. doi: 10.1371/journal.pntd.0002986. eCollection 2014 Jul.
Paracoccidioides lutzii is a new agent of paracoccidioidomycosis (PCM) and has its epicenter localized to the Central-West region of Brazil. Serological diagnosis of PCM caused by P. lutzii has not been established. This study aimed to develop new antigenic preparations from P. lutzii and to apply them in serological techniques to improve the diagnosis of PCM due to P. lutzii. Paracoccidioides lutzii exoantigens, cell free antigen (CFA), and a TCA-precipitated antigen were evaluated in immunodiffusion (ID) tests using a total of 89 patient sera from the Central-West region of Brazil. Seventy-two sera were defined as reactive for P. brasiliensis using traditional antigens (AgPbB339 and gp43). Non-reactive sera for traditional antigens (n = 17) were tested with different P. lutzii preparations and P. lutzii CFA showed 100% reactivity. ELISA was found to be a very useful test to titer anti-P. lutzii antibodies using P. lutzii-CFA preparations. Sera from patients with PCM due to P. lutzii presented with higher antibody titers than PCM due to P. brasiliensis and heterologous sera. In western blot, sera from patients with PCM due to P. lutzii were able to recognize antigenic molecules from the P. lutzii-CFA antigen, but sera from patients with PCM due to P. brasiliensis could not recognize any P. lutzii molecules. Due to the facility of preparing P. lutzii CFA antigens we recommend its use in immunodiffusion tests for the diagnosis of PCM due to P. lutzii. ELISA and western blot can be used as complementary tests.
卢氏副球孢子菌是副球孢子菌病(PCM)的一种新病原体,其中心位于巴西中西部地区。由卢氏副球孢子菌引起的PCM的血清学诊断尚未确立。本研究旨在从卢氏副球孢子菌开发新的抗原制剂,并将其应用于血清学技术,以改善由卢氏副球孢子菌引起的PCM的诊断。使用来自巴西中西部地区的总共89份患者血清,在免疫扩散(ID)试验中评估了卢氏副球孢子菌外抗原、无细胞抗原(CFA)和三氯乙酸沉淀抗原。使用传统抗原(AgPbB339和gp43),72份血清被定义为对巴西副球孢子菌有反应。对传统抗原无反应的血清(n = 17)用不同的卢氏副球孢子菌制剂进行检测,卢氏副球孢子菌CFA显示出100%的反应性。发现ELISA是使用卢氏副球孢子菌-CFA制剂检测抗卢氏副球孢子菌抗体效价的非常有用的试验。由卢氏副球孢子菌引起的PCM患者的血清比由巴西副球孢子菌引起的PCM患者和异源血清呈现出更高的抗体效价。在蛋白质印迹法中,由卢氏副球孢子菌引起的PCM患者的血清能够识别卢氏副球孢子菌-CFA抗原中的抗原分子,但由巴西副球孢子菌引起的PCM患者的血清不能识别任何卢氏副球孢子菌分子。由于制备卢氏副球孢子菌CFA抗原简便,我们建议将其用于免疫扩散试验,以诊断由卢氏副球孢子菌引起的PCM。ELISA和蛋白质印迹法可作为补充试验。