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目的探讨PCR方法诊断泼尼松龙诱导弓形虫隐性感染小鼠复发的敏感性及阿奇霉素对其复发的治疗效果。方法36只20g左右的雌性ICR小鼠以每组6只随机分成正常组(C)、泼尼松龙组(P)、感染弓形虫组(I)、感染弓形虫+阿奇霉素组(IA)、感染弓形虫+泼尼松龙组(IP)、感染弓形虫+泼尼松龙组+阿奇霉素组(IPA)。0周,I组、IA组、IP组和IPA组小鼠分别经腹腔注射感染Prugniaud株弓形虫包囊10个/只;第6～7周,P组、IP组、IPA组小鼠每只每天后肢内侧皮下注射盐酸泼尼松龙1mg,IA组、IPA组小鼠每只每天按250mg/kg剂量腹腔注射阿奇霉素。分别在0、1、2、3、4、5、6、7周对各组小鼠采血,并萃取全血DNA,通过PCR法扩增弓形虫特异性B1基因。第7周剖杀所有小鼠计数其脑组织弓形虫包囊数。结果相比AF14652基因引物,B1基因引物敏感度更高且特异性好;感染弓形虫前5周的小鼠可以通过PCR方法扩增到B1基因特异性产物,5周后PCR法未能扩增出特异性产物;感染弓形虫激素组(IA)小鼠在使用泼尼松龙2周后小鼠全部死亡,感染弓形虫激素治疗组(IP)小鼠死亡率低(P<0.05);相比感染组(I)和感染+阿奇霉素组(IA)和感染+泼尼松龙+阿奇霉素组(IPA),感染+泼尼松龙组(IP)小鼠脑组织包囊个数显著增加(P<0.01)。结论在PCR方法诊断中,B1基因是很好的诊断弓形虫病的目的基因;泼尼松龙能诱导弓形虫隐性感染小鼠的复发并导致小鼠的死亡;阿奇霉素对弓形虫病治疗有效,但不能完全治愈弓形虫病。
Objective To investigate the sensitivity of PCR in the diagnosis of recurrent Toxoplasma gondii infection in mice infected with Toxoplasma gondii and the therapeutic effect of azithromycin on its recurrence. Methods Twenty - six female ICR mice (20g) were randomly divided into normal group (C), prednisolone group (P), Toxoplasma gondii infection group (I), Toxoplasma gondii + azithromycin group (IA) Toxoplasma gondii + prednisone group (IP), Toxoplasma gondii + prednisolone group + azithromycin group (IPA). At 0 week, mice in group I, group IA, group IPA and group IPA were injected intraperitoneally with 10 toxoplasma gondii of Prugniaud respectively. Groups 6, 6 and 7 were divided into group P, IP and IPA Hypothyroid injection of 1 mg of prednisolone hydrochloride was injected into the medial side of the hindlimb respectively. The mice in the IA and IPA groups were injected intraperitoneally with 250 mg / kg azithromycin every day. Blood was collected from all the mice at 0, 1, 2, 3, 4, 5, 6 and 7 weeks, and whole blood DNA was extracted. The Toxoplasma gondii-specific B1 gene was amplified by PCR. At week 7, all mice were sacrificed to count the number of Toxoplasma cysts in their brain tissue. Results The primers of B1 gene were more sensitive and specific than those of AF14652 gene. The mice of the first 5 weeks after infection with Toxoplasma gondii were able to amplify the specific product of B1 gene by PCR. After 5 weeks, the PCR method failed to amplify (IP) mice infected with Toxoplasma gondii had lower mortality (P <0.05). In the mice infected with Toxoplasma gondii (IA) mice, all the mice died after 2 weeks of prednisolone treatment. The numbers of cysts in brain tissue of mice infected with IP +, IP + and IP were significantly higher than those of infected (I) and infected + azithromycin (IA) and infected + prednisolone + azithromycin (IPA) <0.01). Conclusion In the diagnosis of toxoplasmosis, B1 gene is a good gene for the diagnosis of toxoplasmosis. Prednisolone can induce recurrent Toxoplasma gondii infection in mice and cause death in mice. Azithromycin is effective in the treatment of toxoplasmosis , But can not completely cure toxoplasmosis.