title: Comparison of whole blood, serum, and plasma for early detection of candidemia by multiplex-tandem PCR.
abstract: We applied multiplex-tandem PCR (MT-PCR) to 255 EDTA whole-blood specimens, 29 serum specimens, and 24 plasma specimens from 109 patients with Candida bloodstream infection (candidemia) to determine whether a diagnosis could be expedited in comparison with the time to diagnosis by the use of standard blood culture. Overall, the MT-PCR performed better than blood culture with DNA extracted from whole blood from 52/74 (70%) patients, accelerating the time to detection (blood culture flagging) and determination of the pathogenic species (by use of the API 32C system [bioMérieux, Marcy l'Etoile, France]) by up to 4 days (mean, 2.2 days; range, 0.5 to 8 days). Candida DNA was detected more often in serum (71%) and plasma (75%) than in whole blood (54%), although relatively small numbers of serum and plasma specimens were tested. The sensitivity, specificity, positive predictive value, and negative predictive value of the assay with whole blood were 75%, 97%, 95%, and 85%, respectively. Fungal DNA was not detected by MT-PCR in 6/24 (25%) whole-blood samples drawn simultaneously with the positive blood culture sample. MT-PCR performed better with whole-blood specimens stored at -20 degrees C (75%) and when DNA was extracted within 1 week of sampling (66%). The molecular and culture identification results correlated for 61 of 66 patients (92%); one discrepant result was due to misidentification by culture. All but one sample from 53 patients who were at high risk of candidemia but did not have proven disease were negative by MT-PCR. The results demonstrate the good potential of MT-PCR to detect candidemia, to provide Candida species identification prior to blood culture positivity, and to provide improved sensitivity when applied to with serum and plasma specimens.
authors.affiliation: Centre for Infectious Diseases and Microbiology, Institute of Clinical Pathology and Medical Research, Sydney West Area Health Service, Westmead, NSW, Australia.
authors.forename: Anna
authors.initials: A
authors.lastname: Lau
authors.forename: Catriona
authors.initials: C
authors.lastname: Halliday
authors.forename: Sharon C-A
authors.initials: SC
authors.lastname: Chen
authors.forename: E Geoffrey
authors.initials: EG
authors.lastname: Playford
authors.forename: Keith
authors.initials: K
authors.lastname: Stanley
authors.forename: Tania C
authors.initials: TC
authors.lastname: Sorrell
mesh_terms: D001769:Blood / Q000382:microbiology*; D002175:Candida / Q000145:classification / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D002177:Candidiasis / Q000175:diagnosis* / Q000382:microbiology*; D006801:Humans; D010949:Plasma / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D044967:Serum / Q000382:microbiology*; D013997:Time Factors