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[对疑似弓形虫病患者血清学和分子检测结果的回顾性评估]

[Retrospective evaluation of serologic and molecular test results of toxoplasmosis suspected patients].

作者信息

Erdoğan Emrah, Yürük Merve, Sivcan Eda, Karaca Serkan, Temel Hanife, Şabanoğlu Tuba, Gültekin Aslıhan, Şahin İzzet

机构信息

Erciyes University Faculty of Medicine, Department of Parasitology, Kayseri, Turkey.

出版信息

Mikrobiyol Bul. 2019 Jan;53(1):96-105. doi: 10.5578/mb.67469.

DOI:10.5578/mb.67469
PMID:30683043
Abstract

Toxoplasma gondii is a compulsory intracellular protozoan parasite with a wide range of host in warm-blooded vertebrates and has importance in terms of health and economy. Toxoplasmosis is very common because it can infect people with a variety of ways; ingestion of contaminated water and nutrients; raw or undercooked meats containing tissue cysts, blood transfusions, organ transplantantation and transplacental transfer. The aim of this study was to evaluate serologic and molecular test results of toxoplasmosis pre-diagnosed patients. Anti-T.gondii-IgG, anti-T.gondii-IgM ELISA, anti-T.gondii-IgM IFAT and anti-T.gondii-IgG avidity serological tests and PCR tests were applied by using blood, cerebrospinal fluid, amniotic fluid, pericardial fluid and abscess samples from patients who have admitted to Erciyes University Faculty of Medicine Department of Parasitology routine serology and molecular diagnosis laboratories with a pre-diagnosis of toxoplasmosis. Among 6547 patients 3.3% (n= 220) were only IgM positive, 9.2% were both IgG and IgM positive (n= 598). Among male patients, the positivity rates were lower and only IgM seropositive patients were 0.6% (n= 45) while the frequency of both IgG and IgM positive patients was 0.8% (n= 47). The number of both IgG and IgM seropositive cases among new borns, constituting 6.4% (n= 425) of the total number of patients, was 20 (0.3%) and the number of IgM seropositive samples was 25 (0.4%). Only 290 patients positive for IgM antibodies were studied for IFAT and 22 of these patients were positive for anti-T.gondii-IFAT IgM. Anti-T.gondii IgG avidity test was performed in all IgG positive patients regardless of their IgM seropositivity; low avidity was found in 0.7% (n= 18) of IgM-negative patients' sera and equivocal avidity was detected in 6.5% (n= 179). Low avidity was detected in 2.6% of IgM positive patients. Nine of the patients evaluated as anti-T.gondii IgM negative and IgG positive were detected as positive by PCR and two of them were negative. One of these PCR-positive patient's amniotic fluid was sent after the serological test results and detected as positive. Twenty CSF samples were studied by PCR and 7 samples were positive. Also, 8 blood samples which were anti-T.gondii IgM negative and IgG positive were found to bepositive in 7 and negative in one sample with PCR results, subsequently. PCR tests with pericardial fluid and abscess materials were found to be negative. In the case of suspicious or risky situations such as false negatives or false positives resulting from cross-reaction that can occur in ELISA tests, unnecessary medication or interventional approaches can be avoided by applying molecular-based testing at laboratories with appropriate infrastructure. For this reason, we believe that the application of molecular tests in addition to serological tests in risky situations may give more reliable results.

摘要

刚地弓形虫是一种专性细胞内寄生原虫,在温血脊椎动物中有广泛宿主,在健康和经济方面具有重要意义。弓形虫病非常常见,因为它可以通过多种方式感染人类;摄入受污染的水和营养物质;食用含有组织包囊的生肉或未煮熟的肉类、输血、器官移植和经胎盘传播。本研究的目的是评估弓形虫病预诊断患者的血清学和分子检测结果。对埃尔西耶斯大学医学院寄生虫学系常规血清学和分子诊断实验室收治的、预诊断为弓形虫病的患者,采集血液、脑脊液、羊水、心包液和脓肿样本,进行抗刚地弓形虫IgG、抗刚地弓形虫IgM ELISA、抗刚地弓形虫IgM间接荧光抗体试验(IFAT)、抗刚地弓形虫IgG亲和力血清学检测及PCR检测。在6547例患者中,3.3%(n = 220)仅IgM阳性,9.2% IgG和IgM均阳性(n = 598)。男性患者中,阳性率较低,仅IgM血清阳性患者为0.6%(n = 45),而IgG和IgM均阳性患者的频率为0.8%(n = 47)。新生儿中IgG和IgM均血清阳性的病例数占患者总数的6.4%(n = 425),为20例(0.3%),IgM血清阳性样本数为25例(0.4%)。仅对290例IgM抗体阳性患者进行了IFAT检测,其中22例抗刚地弓形虫IFAT IgM阳性。对所有IgG阳性患者均进行了抗刚地弓形虫IgG亲和力检测,无论其IgM血清阳性情况如何;IgM阴性患者血清中0.7%(n = 18)检测到低亲和力,6.5%(n = 179)检测到可疑亲和力。IgM阳性患者中2.6%检测到低亲和力。9例被评估为抗刚地弓形虫IgM阴性且IgG阳性的患者经PCR检测为阳性,其中2例为阴性。其中1例PCR阳性患者的羊水在血清学检测结果出来后送检,检测为阳性。对20份脑脊液样本进行了PCR检测,7份样本呈阳性。此外,8份抗刚地弓形虫IgM阴性且IgG阳性的血液样本经PCR检测,7份呈阳性,1份呈阴性。心包液和脓肿样本的PCR检测结果均为阴性。在ELISA检测可能出现交叉反应导致假阴性或假阳性等可疑或危险情况时,通过在具备适当基础设施的实验室应用基于分子的检测,可以避免不必要的药物治疗或干预措施。因此,我们认为在危险情况下除血清学检测外应用分子检测可能会得到更可靠的结果。

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