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초록
Background
Cytomegalovirus (CMV) infection is a major cause of morbidity and mortality in immunoco-mpromised patients. We compared the abilities of the recently developed Real-Q Cytomegalovirus Kit (Bio-sewoom Inc., Korea) and the previously used PANA mPCR CMV Detection Kit (Panagene Inc., Korea) to detect CMV.
Methods
We analyzed 300 samples (whole blood: 262, urine: 37, CSF: 1) submitted for qualitative CMV PCR testing during October 2011 at Yonsei University College of Medicine Severance Hospital. real-time PCR was performed with a Real-Q Cytomegalovirus Kit and conventional PCR was conducted with a PANAmPCR CMV Detection Kit.
Results
The positive rates of both real-time PCR and conventional PCR were 25.3% (76/300), and the kappa coefficient (K) was 0.96 (95% confidence interval (CI), 0.93-1.00). The concordance rate of the Real-Q Cytomegalovirus Kit and the PANAmPCR CMV Detection Kit was 98.7% (296/300), and four out of 300 samples showed discordant results. If the concordant results of 296 samples and the four results confirmed by direct sequencing were assumed to be true, the sensitivity and specificity of the Real-Q Cytomegalovirus Kit were 97.4% (95% CI, 93.8-100.0%) and 99.1% (95% CI, 97.9-100.0%), respectively.
Conclusion
The recently developed Real-Q Cytome-galovirus Kit showed excellent sensitivity and specificity, and had a high concordance rate with the previously established PANAmPCR CMV Detection Kit, which uses conventional PCR. Furthermore, real-time PCR could decrease the test time, as the electro-phoresis step required for conventional PCR is not required for real-time PCR.
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Table 1.
Demographic characteristics of the patients
| Characteristics | No. of patients | Percentage (%) |
|---|---|---|
| Sex | ||
| Male | 168 | 56.0 |
| Female | 132 | 44.0 |
| Age (years) | ||
| Under 20 | 48 | 16.0 |
| 20-30 | 26 | 8.7 |
| 30-40 | 42 | 14.0 |
| 40-50 | 40 | 13.3 |
| 50-60 | 73 | 24.3 |
| 60 and over | 71 | 23.7 |
| Ward | ||
| ICU | 37 | 12.3 |
| General | 263 | 87.7 |
| Diagnosis | ||
| Hematologic disease | 175 | 58.3 |
| Solid cancer | 13 | 4.3 |
| Renal disease | 23 | 7.7 |
| Other∗ | 89 | 29.7 |
| Specimen type | ||
| Whold blood | 262 | 87.3 |
| Urine | 37 | 12.3 |
| CSF | 1 | 0.3 |
Table 2.
Comparison of real-time PCR and conventional PCR by kappa agreement and McNemar test for detection of CMV infection
| Parameters | Real-time PCR (n=300) | ||
|---|---|---|---|
| Positive | Negative | ||
| Conventional PCR | Positive | 74 | 2 |
| Negative | 2 | 222 | |
| Kappa agreement∗ | 0.96 (0.93 to 1.00) | ||
| P value of McNemar test | 1.0000 | ||
| Agreement (%) | 98.7 | ||
Table 3.
Results of CMV antibodies on the discrepancy (n=4) between real-time PCR and conventional PCR
| Sample No. | Real-time | Ct of real-time | Conventional | CMV IgM | CMV IgG | Direct sequencing∗ | ∗ Diagnosis | CMV associated diagnosis∗ |
|---|---|---|---|---|---|---|---|---|
| 1 | Negative | -† | Positive | Negative | Positive | Positive | Hepatocellular carcinoma | Leukopenia, thrombocytopenia |
| 2 | Positive | 39.87 | Negative | Negative | Positive | -‡ | Chronic myelocytic leukemia | Leukopenia, thrombocytopenia, enteritis |
| 3 | Negative | -† | Positive | Negative | Positive | Positive | Acute lymphocytic leukemia | Leukopenia, thrombocytopenia, retinitis |
| 4 | Positive | 39.91 | Negative | Not tested | Not tested | -‡ | Hodgkin's disease | Leukopenia, thrombocytopenia |
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