The role of a real-time PCR technology for rapid detection and identification of bacterial and fungal pathogens in whole-blood samples.
The rapid diagnosis of pathogens and prompt initiation of appropriate antibiotic therapy are critical factors to reduce the morbidity and mortality associated with sepsis. In this study, we evaluated a multiplex polymerase chain reaction (PCR-M) test that detects bacteria and fungi in whole-blood specimens, comparing its features to those of a blood culture (BC). Following evaluation of the performance for sensitivity and specificity of PCR-M, 78 blood samples from 54 patients with suspected bacterial infections were evaluated. Whole-blood samples for PCR-M were collected at the same time as BC, and PCR-M results were compared with BC results. As a result, minimum sensitivity of the kit was 1-100 cfu/ml. The PCR-M test correctly identified specificity for 13 out of 14 strains blinded to the assay analyst. Of 78 blood samples examined, 56 (72%) were negative by both methods, and 22 (28%) were positive by at least one of the two methods. PCR-M detected organisms in 21 cases (27%) compared with 12 cases (15%) in BC. The correlation of positives between PCR-M and BC was 92% (11/12), and both methods identified the same organisms in these 11 cases. With higher positive rate compared with BC, PCR-M could detect and identify potentially significant microorganisms within a few hours by using a small volume of a single whole-blood sample. Early detection of microorganisms has the potential to facilitate early determination of appropriate treatment and antimicrobial selection.
Department of Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan. obara@sc.itc.keio.ac.jp
Hideaki
H
Obara
Naoki
N
Aikawa
Naoki
N
Hasegawa
Shingo
S
Hori
Yasuo
Y
Ikeda
Yoshio
Y
Kobayashi
Mitsuru
M
Murata
Shinichiro
S
Okamoto
Junzo
J
Takeda
Minoru
M
Tanabe
Yasuhiko
Y
Sakakura
Hiroyuki
H
Ginba
Masaki
M
Kitajima
Yuko
Y
Kitagawa
D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000097:blood*; D001769:Blood / Q000382:microbiology*; D005260:Female; D005658:Fungi / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009181:Mycoses / Q000097:blood*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D018805:Sepsis / Q000097:blood / Q000382:microbiology; D018746:Systemic Inflammatory Response Syndrome / Q000097:blood / Q000382:microbiology