title: Bacterial rRNA-targeted reverse transcription-PCR used to identify pathogens responsible for fever with neutropenia.
abstract: The purpose of this study was to evaluate the clinical utility of bacterial rRNA-targeted reverse transcription-quantitative PCR (BrRNA RT-qPCR) assays for identifying the bacterial pathogens that cause fever with neutropenia in pediatric cancer patients, by comparing the bacterial detection rate of this technique with that of blood culture. One milliliter of blood was collected from pediatric patients who developed fever with neutropenia following cancer chemotherapy. BrRNA RT-qPCR was performed using 16 primer sets, each designed for a specific type of bacteria. The entire BrRNA RT-qPCR procedure took less than 5 h. Blood culture was performed at the same time, following the standard institutional procedure. Blood from 13 patients was collected during 23 febrile neutropenic episodes. Of these samples, bacteria were identified in 16 by BrRNA RT-qPCR (69.6%) and in 4 by blood culture (17.4%, P<0.001). In all 4 blood culture-positive samples, BrRNA RT-qPCR detected the same type of bacteria as that identified by culture. In 9 samples, more than 4 types of bacteria were identified simultaneously by BrRNA RT-qPCR, most of which were anaerobic bacteria known to be part of the gut flora. We conclude that BrRNA RT-qPCR could be useful in the diagnosis of fever with neutropenia, given its high bacterial detection rate, short turnaround time, and the small blood sample required compared with the standard blood culture techniques. Our findings also indicate that anaerobic intestinal bacteria, which are difficult to detect by standard culture techniques, may be responsible for some cases of febrile neutropenia.
authors.affiliation: Department of Pediatrics and Adolescent Medicine, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan.
authors.forename: Sachi
authors.initials: S
authors.lastname: Sakaguchi
authors.forename: Masahiro
authors.initials: M
authors.lastname: Saito
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Tsuji
authors.forename: Takashi
authors.initials: T
authors.lastname: Asahara
authors.forename: Oto
authors.initials: O
authors.lastname: Takata
authors.forename: Junya
authors.initials: J
authors.lastname: Fujimura
authors.forename: Satoru
authors.initials: S
authors.lastname: Nagata
authors.forename: Koji
authors.initials: K
authors.lastname: Nomoto
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Shimizu
mesh_terms: D000293:Adolescent; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D001424:Bacterial Infections / Q000175:diagnosis* / Q000382:microbiology; D001431:Bacteriological Techniques / Q000379:methods*; D001769:Blood / Q000382:microbiology; D002648:Child; D002675:Child, Preschool; D017931:DNA Primers / Q000235:genetics; D005260:Female; D005335:Fever of Unknown Origin / Q000150:complications / Q000382:microbiology*; D006801:Humans; D016867:Immunocompromised Host; D007223:Infant; D008297:Male; D009369:Neoplasms / Q000150:complications / Q000188:drug therapy; D009503:Neutropenia / Q000150:complications / Q000382:microbiology*; D012329:RNA, Bacterial / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors
chemical_list: D017931:DNA Primers; D012329:RNA, Bacterial; D012335:RNA, Ribosomal