Rapid and sensitive diagnosis of cytomegalovirus and Pneumocystis carinii pneumonia in patients with haematological neoplasia by using capillary polymerase chain reaction.
We attempted the simultaneous detection of cytomegalovirus DNA (CMV-DNA) and Pneumocystis carinii (carinii-DNA) in sputum samples obtained from 20 patients with haematological neoplasm with pneumonia, using rapid cycle DNA amplification (capillary PCR). We used a thermal cycler for capillary PCR which featured recirculation of hot air for rapid temperature control of 10 microliters reaction samples in thin glass capillary tubes. We extracted DNA from patients' sputa using a simple method. A comparison of the results obtained using the phenol extraction-ethanol precipitation method and those obtained using our simple method was made, and demonstrated complete agreement between the two. For detection of CMV-DNA and P. carinii-DNA with capillary PCR it was not necessary to vary temperature setting based on the primers used. Therefore, capillary PCR was used for the simultaneous detection of CMV and P. carinii. After amplification, the total time required for which was 20 min, amplified products were electrophoresed on agarose gels and visualized with ethidium bromide. Product sensitivity was higher with capillary PCR than with conventional PCR. We conclude that capillary PCR amplification is a valuable tool for rapid and simple diagnosis of CMV and P. carinii pneumonias.
First Department of Internal Medicine, Kurume University, Fukuoka, Japan.
J
J
Honda
T
T
Hoshino
H
H
Natori
S
S
Tokisawa
H
H
Akiyoshi
S
S
Nakahara
K
K
Oizumi
D000328:Adult; D000368:Aged; D000970:Antineoplastic Agents / Q000009:adverse effects; D001483:Base Sequence; D003586:Cytomegalovirus Infections / Q000175:diagnosis*; D004271:DNA, Fungal / Q000737:chemistry; D004279:DNA, Viral / Q000737:chemistry; D004587:Electrophoresis, Agar Gel; D005260:Female; D006801:Humans; D007938:Leukemia / Q000382:microbiology; D008223:Lymphoma / Q000382:microbiology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009894:Opportunistic Infections / Q000175:diagnosis*; D011020:Pneumonia, Pneumocystis / Q000175:diagnosis*; D011024:Pneumonia, Viral / Q000175:diagnosis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
D000970:Antineoplastic Agents; D004271:DNA, Fungal; D004279:DNA, Viral