Comparison of a conventional polymerase chain reaction with real-time polymerase chain reaction for the detection of neurotropic viruses in cerebrospinal fluid samples

被引:16
作者
Ramamurthy, M. [1 ]
Alexander, M. [2 ]
Aaron, S. [2 ]
Kannangai, R. [1 ]
Ravi, V. [3 ]
Sridharan, G. [1 ]
Abraham, A. M. [1 ]
机构
[1] Christian Med Coll & Hosp, Dept Clin Virol, Vellore 632004, Tamil Nadu, India
[2] Christian Med Coll & Hosp, Dept Neurol, Vellore 632004, Tamil Nadu, India
[3] Natl Inst Mental Hlth & Neurosci, Dept Neurovirol, Bangalore 560029, Karnataka, India
关键词
CNS infections; neurotropic viruses; real-time PCR; CENTRAL-NERVOUS-SYSTEM; EPSTEIN-BARR-VIRUS; PCR ASSAY; HERPESVIRUS INFECTIONS; VARICELLA-ZOSTER; NESTED PCR; DNA; QUANTIFICATION; SIMPLEX;
D O I
10.4103/0255-0857.81777
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Purpose: To compare a conventional polymerase chain reaction (PCR) and real-time PCR for the detection of neurotropic DNA viruses. Materials and Methods: A total of 147 cerebrospinal fl uid (CSF) samples was collected from patients attending a tertiary care hospital in South India for a period from 2005 to 2008. All these samples were tested using a conventional multiplex/uniplex PCR and a real-time multiplex/uniplex PCR. This technique was used to detect a large number of herpes viruses responsible for central nervous system infections, including HSV-1, HSV-2, VZV, CMV and EBV and the polyoma virus JCV. Results: Overall, in the entire set of samples, the real-time PCR yielded 88 (59.9%) positives and conventional PCR had six (4.1%) positives. Conclusion: Our results suggest that the real-time PCR assay was more sensitive compared with the conventional PCR. The advantage of real-time PCR is that it can be performed much faster than conventional PCR. Real-time PCR is less time-consuming, less labour-intensive and also reduces the chance of contamination as there is no post-amplifi cation procedure. In the entire study population, the major viruses detected using real-time PCR were EBV (34%), HSV-2 (10.8%) and VZV (6.8%).
引用
收藏
页码:102 / 109
页数:8
相关论文
共 19 条
[1]   Rapid quantification and differentiation of human polyomavirus DNA in undiluted urine from patients after bone marrow transplantation [J].
Biel, SS ;
Held, TK ;
Landt, O ;
Niedrig, M ;
Gelderblom, HR ;
Siegert, W ;
Nitsche, P .
JOURNAL OF CLINICAL MICROBIOLOGY, 2000, 38 (10) :3689-3695
[2]  
Cassady Kevin A., 2002, P27
[3]   Comparison of nested PCR and real time PCR of herpesvirus infections of central nervous system in HIV patients [J].
Drago, L ;
Lombardi, A ;
De Vecchi, E ;
Giuliani, G ;
Bartolone, R ;
Gismondo, MR .
BMC INFECTIOUS DISEASES, 2004, 4 (1)
[4]  
Gubler D.J., 2007, FIELDS VIROLOGY, V5th, P1153
[5]   Practical experience of high throughput real time PCR in the routine diagnostic virology setting [J].
Gunson, R. N. ;
Collins, T. C. ;
Carman, W. F. .
JOURNAL OF CLINICAL VIROLOGY, 2006, 35 (04) :355-367
[6]   New PCR systems to confirm real-time PCR detection of Mycobacterium avium subsp paratuberculosis [J].
Herthnek, David ;
Bolske, Goran .
BMC MICROBIOLOGY, 2006, 6 (1)
[7]  
JAWETZ, 2004, MED MICROBIOLOGY, P429
[8]   A comparison study of different PCR assays in measuring circulating plasma Epstein-Barr virus DNA levels in patients with nasopharyngeal carcinoma [J].
Le, QT ;
Jones, CD ;
Yau, TK ;
Shirazi, HA ;
Wong, PH ;
Thomas, EN ;
Patterson, BK ;
Lee, AWM ;
Zehnder, JL .
CLINICAL CANCER RESEARCH, 2005, 11 (16) :5700-5707
[9]   Role of cell culture for virus detection in the age of technology [J].
Leland, Diane S. ;
Ginocchio, Christine C. .
CLINICAL MICROBIOLOGY REVIEWS, 2007, 20 (01) :49-+
[10]   Laboratory diagnosis of common herpesvirus infections of the central nervous system by a multiplex PCR assay [J].
Markoulatos, P ;
Georgopoulou, A ;
Siafakas, N ;
Plakokefalos, E ;
Tzanakaki, G ;
Kourea-Kremastinou, J .
JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (12) :4426-4432