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首页> 外文期刊>Journal of Clinical and Diagnostic Research >Diagnosis of Acute Q Fever by Detection of Coxiella burnetii DNA using Real-Time PCR, Employing a Commercial Genesig Easy Kit DC10-DC13
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Diagnosis of Acute Q Fever by Detection of Coxiella burnetii DNA using Real-Time PCR, Employing a Commercial Genesig Easy Kit DC10-DC13

机译：使用实时Genesig Easy Kit DC10-DC13，通过实时PCR检测柯氏氏杆菌DNA诊断急性Q发热

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Introduction: Query (Q) fever is an important zoonosis and a cause of concern for humans, due to the potential bioterrorism threat posed by the causative agent, Coxiella burnetii. Because of the danger of contracting the illness, isolation attempts are seldom made. Serological and molecular diagnostic tests are the main option.Aim: To study the prevalence of acute Q fever in Puducherry and surrounding districts of Tamil Nadu, India, employing a new commercial Real-Time Polymerase Chain Reaction (PCR) kit and confirming it by the gold standard Immunofluorescence Assay (IFA).Materials and Methods: Acute phase blood samples from 72 consecutive febrile patients and 24 healthy individuals were included in this prospective study. DNA was extracted from the buffy coats and preserved at -80°C. Detection of C. burnetii was carried out employing a commercial Real-Time PCR kit. Serum samples were tested for IgM (Phase I+II) and IgG (Phase I+II) by QM-120 and QG-120, Coxiella burnetii IFA Fuller Laboratories, California, USA. Sensitivity, Specificity, Positive Predictive Value (PPV) and Negative Predictive Value (NPV) were calculated keeping IFA as the reference.Results: Presumptive diagnosis of acute Q fever was made in two febrile patients by the Genesig Easy kit (2.78%). In addition to these two PCR positive cases, one more patient was positive for both Phase II IgM and Phase II IgG antibodies by the gold standard IFA. All 24 healthy controls were negative for Q fever by both PCR and IFA. The sensitivity, specificity, NPV and PPV for Genesig Easy kit PCR were: 66.67%, 100%, 100% and 98.57 % respectively against IFA as the reference.Conclusion: The true prevalence of Q fever in India and other developing countries is poorly understood, owing to the difficulties in the diagnosis of this infection. Since molecular diagnostic tests have good specificity and are mandated for confirmation of single acute samples, validation of commercial Q fever PCR kits is the need of the hour. Genesig Easy kit in our hands was found to be reliable with the moderate sensitivity and high specificity. Performing both PCR (with acute specimens) and IFA (with paired sera) would be ideal for Q fever diagnosis.
机译：简介：查询（Q）发烧是一种重要的人畜共患病，也是人类关注的原因，这是由于病原体柯氏杆菌引起的潜在的生物恐怖主义威胁。由于感染疾病的危险，很少进行隔离尝试。目的：通过新型商业实时聚合酶链反应（PCR）试剂​​盒并通过验证，以研究印度泰米尔纳德邦Puducherry及其周边地区的急性Q发热的患病率。金标准免疫荧光测定法（IFA）。材料与方法：前瞻性研究包括来自72位连续发热患者和24位健康个体的急性期血样。从血沉棕黄层中提取DNA，并保存在-80°C。伯氏梭状芽胞杆菌的检测是使用商业实时PCR试剂盒进行的。通过QM-120和QG-120（美国加利福尼亚州柯尼氏杆菌IFA富勒实验室）测试了血清样品的IgM（I + II期）和IgG（I + II期）。以IFA为参考，计算敏感性，特异性，阳性预测值（PPV）和阴性预测值（NPV）。结果：通过Genesig Easy试剂盒（2.78％）对两名高热患者进行了急性Q热的推测诊断。除了这两个PCR阳性病例外，又有一个金标准IFA的II期IgM和II期IgG抗体均为阳性的患者。通过PCR和IFA，所有24名健康对照组的Q发热均为阴性。 Genesig Easy试剂盒PCR的敏感性，特异性，NPV和PPV分别为：以IFA为参考分别为：66.67％，100％，100％和98.57％。 ，由于这种感染的诊断困难。由于分子诊断测试具有良好的特异性，并且要求确认单个急性样品，因此需要一小时的时间来验证商业Q发烧PCR试剂盒。发现我们手中的Genesig Easy试剂盒具有中等灵敏度和高特异性，是可靠的。同时进行PCR（对急性标本）和IFA（对与血清配对）对于Q热诊断是理想的。

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