Detection of late pp67-mRNA by NASBA in peripheral blood for the diagnosis of human cytomegalovirus disease in AIDS patients.
OBJECTIVE
To evaluate the value of late-pp67-mRNA nucleic acid sequence-based amplification (NASBA) in comparison to DNA-PCR, blood culture and pp65-antigenemia assay for the detection of human cytomegalovirus (HCMV) disease in HIV-infected patients.


METHODS
The results of pp67-mRNA NASBA, DNA-PCR, culture and pp65-antigenemia assay were compared in 402 whole blood specimens of 98 HIV-infected patients with a low CD4 lymphocyte count who had not yet received highly active antiretroviral therapy (HAART). Thirty-seven samples were obtained from 30 patients with a diagnosis of HCMV disease and 365 samples from 68 patients without HCMV disease.


RESULTS
The highest agreement of test results was observed between pp67-mRNA NASBA and quantitative pp65-antigenemia, with a threshold of nine antigen-positive cells/10(5) leukocytes (kappa-value 0.70, 95% CI=0.58-0.82). The sensitivity of pp67-mRNA NASBA for the diagnosis of HCMV disease (59.3%) was identical to that of the quantitative pp65-antigenemia assay, higher than that of the blood culture (48.2%) but lower than that of the DNA-PCR (77.8%). Pp67-mRNA NASBA (92.3%), quantitative pp65-antigenemia assay (92.3%) and blood culture (93.9%) were highly specific for the diagnosis of HCMV disease and as a result, had a higher positive predictive value (76.2, 76.2 and 76.5%, respectively) than the qualitative DNA-PCR (58.3%) and the qualitative pp65-antigenemia assay (47.6%).


CONCLUSIONS
pp67-mRNA NASBA, an easy and rapid to perform assay, well-standardised by virtue of co-amplified internal system control RNA, provides a high specificity and positive predictive value for the diagnosis of HCMV disease in HIV-infected patients, comparable to that of the quantitative pp65-antigenemia assay and blood culture.
Department of Internal Medicine, National AIDS Therapy Evaluation Centre, Academic Medical Centre, University of Amsterdam, Bouwdeel T, Meibergdreef 9, 1100 DD, Amsterdam, The Netherlands. s.n.blank@amc.uva.nl
B S N
BS
Blank
P L
PL
Meenhorst
W
W
Pauw
J W
JW
Mulder
W C
WC
van Dijk
P H M
PH
Smits
F
F
Roeles
J M
JM
Middeldorp
J M A
JM
Lange
D017088:AIDS-Related Opportunistic Infections / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D000328:Adult; D000956:Antigens, Viral / Q000097:blood / Q000235:genetics*; D023241:Antiretroviral Therapy, Highly Active; D003587:Cytomegalovirus / Q000235:genetics; D003586:Cytomegalovirus Infections / Q000097:blood / Q000175:diagnosis / Q000821:virology*; D005260:Female; D005500:Follow-Up Studies; D005784:Gene Amplification / Q000235:genetics; D015658:HIV Infections / Q000150:complications / Q000188:drug therapy; D006801:Humans; D008297:Male; D010750:Phosphoproteins / Q000097:blood / Q000235:genetics*; D011446:Prospective Studies; D012333:RNA, Messenger / Q000097:blood*; D012367:RNA, Viral / Q000097:blood*; D012680:Sensitivity and Specificity; D014763:Viral Matrix Proteins / Q000097:blood / Q000235:genetics*
D000956:Antigens, Viral; D010750:Phosphoproteins; D012333:RNA, Messenger; D012367:RNA, Viral; D014763:Viral Matrix Proteins; C464963:pp67 protein, human cytomegalovirus