TABLE 15.

Detection of enterovirus cDNA by real-time PCR

ReferenceCultureReal-time PCRComments
No. of specimensNo. positive (%)No. of specimensNo. positive (%)Test platform/ gene target
Verstrepen et al. 2001 (515)7017 (24.3)7019 (27.1)ABI Prism/5′ nontranslated regionSensitivity of real-time PCR was 100% compared with cell culture.
Read et al. 2001 (403)50 (originally found positive by conventional PCR)49 (98)LightCycler/5′ nontranslated regionPublication described conversion of a conventional multiplex PCR assay that detects HSV-1, HSV-2, VZV, and enterovirus with the LightCycler system.
Corless et al. 2002 (78)200 (97, CSF; 103, throat swabs) giving previously negative results in cell culture97 (CSF), 103 (throat swabs)An additional 33 (15.9) enterovirus and 2 (1) parechoviruses (formerly echoviruses 22 and 23) identified by PCRABI Prism/5′ nontranslated regionReal-time PCR was 11.5% more sensitive than cell cultures for the diagnosis of enterovirus infections using CSF specimens. Based on limiting dilutions, the TaqMan enterovirus and parechovirus PCR showed an increase of two orders of magnitude compared to cell culture with a sensitivity of 100% when assessed using enterovirus cell culture-positive samples.
Watkins-Riedel et al. 2002 (528)60 (stool, 12; CSF, 38; serum, 8; throat swabs, 2)6/12 feces (50), 2/38 CSF (5.3)606/12 feces (50), 2/38 CSF (5.3)LightCycler 5′ nontranslated regionSensitivity of the real-time PCR was 10-100-fold higher compared to AMPLICOR EV test.
Monpoeho et al. 2002 (319)104 (CSF)22 (21.2)104 (CSF)61 (58.7)ABI Prism/5′ nontranslated regionReal-time PCR allows a large number of samples to be screened rapidly during an epidemic and its sensitivity, simplicity, and reproducibility make it a highly reliable and suitable tool in the clinical laboratory.
Nijhuis et al. 2002 (344)41 (feces), 8 (CSF), 43 (bronchoalveolar lavage)9 (22.0), 3 (37.5), 041 (feces), 8 (CSF), 43 (bronchoalveolar lavage)10 (24.4), 4 (50.0), 2 (4.7)ABI Prism/5′ nontranslated regionReal-time assay was robust and easily standardized, which make it an excellent alternative for conventional time-consuming viral culture.
Rabenau et al. 2002 (390)109 (CSF)23 (21)LightCycler/5′ nontranslated regionPerformance characteristics of real-time PCR were comparable to those using a conventional PCR assay. Compared with the conventional laboratory-developed assay real-time PCR was less labor intensive and easy to use.
Kares et al. 2003 (207)3212 (37.5)55 (CSF, 21; stool, 32; nasopharyngeal aspirate, 3)32 (58.1)LightCycler/5′ nontranslated region, SYBR Green I dye detectionReal-time assays were of equal sensitivity to laboratory developed developed real-time PCR test. Fifteen of 20 samples which were negative in cell culture were positive by real-time PCR.
Verboon-Maciolek et al. 2003 (514)19 (CSF)4 (21.1)19 (CSF and serum)5 CSF (26.3), 9 serum (47.4)ABI Prism 5′ nontranslated regionStudy population was infants ≤60 days old who received a clinical diagnosis of sepsis. Enterovirus infections are an important cause of sepsis in infants admitted to the hospital.