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三级护理医院中疑似新型隐球菌脑膜炎的HIV阳性和阴性患者脑脊液样本中抗原检测与巢式PCR的比较

Comparison of Antigen Detection and Nested PCR in CSF Samples of HIV Positive and Negative Patients with Suspected Cryptococcal Meningitis in a Tertiary Care Hospital.

作者信息

Kumari Sunita, Verma Rajesh Kumar, Singh Dharmendra Prasad, Yadav Ramakant

机构信息

Lecturer, Department of Microbiology, UPRIMS & R , Saifai, Etawah, (UP), India .

Associate Professor, Department of Microbiology, UPRIMS & R , Saifai, Etawah, (UP), India .

出版信息

J Clin Diagn Res. 2016 Apr;10(4):DC12-5. doi: 10.7860/JCDR/2016/18322.7619. Epub 2016 Apr 1.

Abstract

INTRODUCTION

The cases of cryptococcal meningitis and other forms of cryptococcosis have increased in recent time and the present scenario of the condition with significant morbidity and mortality is actually posing a serious threat to the community, so an early and prompt diagnosis is necessary to prevent serious complications and thus improving the overall disease outcome.

AIM

Comparison of diagnostic efficacy of nested Polymerase Chain Reaction (PCR) with Latex Agglutination Test (LAT) in the Cerebro Spinal Fluid (CSF) samples of the cases of meningitis in HIV positive and negative cases.

MATERIALS AND METHODS

We have compared the diagnostic efficacy of Latex Agglutination Test (LAT) with nested Polymerase Chain Reaction (PCR) in 200 Cerebrospinal Fluid (CSF) samples, including 14 HIV positive also, in the cases of suspected cryptococcal meningitis. Nested PCR was done in all cases reporting positive by LAT and results were then compared with that of India ink and culture on Sabouraud Dextrose Agar (SDA), and the isolates were further identified by urease, nitrate and sugar assimilation tests.

RESULTS

Of the 200 cases, including 14 HIV positive, LAT was positive in 46 cases while 154 were negative. Out of these 46 LAT positive cases, nested PCR was positive in 40 cases only, while culture and India ink was positive in 38 and 33 cases respectively. Majority of the cases, 30 (65.2%) were between age group 21-50 years, while 2 (4.3%) in 0-20, and 14 (30.4%) in 51-80 years age group.

CONCLUSION

Although negative staining like India ink and nigrosin are most widely used techniques, but these suffer with subjective error. Rapid method like LAT is available but it always has the scope of false positive and negative results. In such cases nested PCR can help in establishing final diagnosis.

摘要

引言

近年来,隐球菌性脑膜炎及其他形式的隐球菌病病例有所增加,当前这种具有显著发病率和死亡率的疾病状况实际上对社区构成了严重威胁,因此早期及时诊断对于预防严重并发症并改善整体疾病结局至关重要。

目的

比较巢式聚合酶链反应(PCR)与乳胶凝集试验(LAT)在HIV阳性和阴性脑膜炎病例脑脊液(CSF)样本中的诊断效能。

材料与方法

我们比较了乳胶凝集试验(LAT)与巢式聚合酶链反应(PCR)在200例疑似隐球菌性脑膜炎脑脊液(CSF)样本中的诊断效能,其中包括14例HIV阳性样本。对所有LAT检测呈阳性的病例进行巢式PCR检测,然后将结果与印度墨汁染色及在沙氏葡萄糖琼脂(SDA)上的培养结果进行比较,并通过脲酶、硝酸盐和糖同化试验进一步鉴定分离株。

结果

在这200例病例中,包括14例HIV阳性病例,LAT检测呈阳性的有46例,阴性的有154例。在这46例LAT阳性病例中,巢式PCR仅在40例中呈阳性,而培养和印度墨汁染色分别在38例和33例中呈阳性。大多数病例(30例,65.2%)年龄在21 - 50岁之间,0 - 20岁的有2例(4.3%),51 - 80岁年龄组的有14例(30.4%)。

结论

尽管像印度墨汁和黑色素负染这样的方法是最广泛使用的技术,但这些方法存在主观误差。像LAT这样的快速方法虽然可用,但始终存在假阳性和假阴性结果的可能性。在这种情况下,巢式PCR有助于确立最终诊断。

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本文引用的文献

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Neurocryptococcosis: diagnosis by PCR method.神经隐球菌病:通过聚合酶链反应(PCR)方法进行诊断
Rev Inst Med Trop Sao Paulo. 2004 Jul-Aug;46(4):203-7. doi: 10.1590/s0036-46652004000400006. Epub 2004 Sep 3.
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Cryptococcosis.隐球菌病
Infect Dis Clin North Am. 2002 Dec;16(4):837-74, v-vi. doi: 10.1016/s0891-5520(02)00036-3.

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