Comparison of the performance of polymerase chain reaction and pp65 antigenemia for the detection of human cytomegalovirus in immunosuppressed patients

Abstract

Comparison of the performance of polymerase chain reaction and pp65 antigenemia for the detection of human cytomegalovirus in immunosuppressed patientsComparação do desempenho da reação em cadeia da polimerase e antigenemia pp65 para detecção de citomegalovírus humano em pacientes imunossuprimidos ABSTRACT Introduction: Human cytomegalovirus (HCMV) is often reactive in latently infected immunosuppressed patients. Accordingly, HCMV remains one of the most common infections following solid organ and hemopoietic stem cell transplantations, resulting in significant morbidity, graft loss and occasional mortality. The early diagnosis of HCMV disease is important in immunosuppressed patients, since in these individuals, preemptive treatment is useful. The objective of this study was to compare the performance of the in-house qualitative polymerase chain reaction (PCR) and pp65 antigenemia to HCMV infection in immunosuppressed patients in the Hospital de Clínicas of Porto Alegre (HCPA). Methods: A total of 216 blood samples collected between August 2006 and January 2007 were investigated. Results: Among the samples analyzed, 81 (37.5%) were HCMV-positive by PCR, while 48 (22.2%) were positive for antigenemia. Considering antigenemia as the gold standard, sensitivity, specificity, positive predictive values and negative predictive values for PCR were 87.5%, 76.8%, 51.8% and 95.5% respectively. Conclusions: These results demonstrated that qualitative PCR has high sensitivity and negative predictive value (NPV). Consequently PCR is especially indicated for the initial diagnosis of HCMV infection. In the case of preemptive treatment strategy, identification of patients at high-risk for HCMV disease is fundamental and PCR can be useful tool.

Cytomegalovirus Identification in Blood and Urine of Newborns by Nested PCR

“…Polymerase chain reaction technique offers speed and high sensitivity based on selective amplification of specific nucleic acid sequences from small amounts of biological specimens. Furthermore, the storage convenience of target specimens, such as its freezing at -20°C or -80°C, prior to test, gives PCR a larger advantage over any other technique, due to the stability of nucleic acids (5,8).…”

“…Polymerase chain reaction was carried out and DNA amplification was performed using nested PCR method, using the outside primers (5'-TG AGG AAT GTC TTC AGC-3'and 5'-TC TCC TCG AGG ATG AGA-3') in the first reaction, which yields an amplicon of 347 bp, serving as template for the second reaction which used the internal primers (5'-CCA ACT GCC AGA TCT TCA T-3' and 5'-TCG AGA TCC CCC AGG TTG TA-3') for an amplicon of 297 bp, according to Martiny et al (5).…”