title: Melting Temperature Mapping Method: A Novel Method for Rapid Identification of Unknown Pathogenic Microorganisms within Three Hours of Sample Collection.
abstract: Acquiring the earliest possible identification of pathogenic microorganisms is critical for selecting the appropriate antimicrobial therapy in infected patients. We herein report the novel "melting temperature (Tm) mapping method" for rapidly identifying the dominant bacteria in a clinical sample from sterile sites. Employing only seven primer sets, more than 100 bacterial species can be identified. In particular, using the Difference Value, it is possible to identify samples suitable for Tm mapping identification. Moreover, this method can be used to rapidly diagnose the absence of bacteria in clinical samples. We tested the Tm mapping method using 200 whole blood samples obtained from patients with suspected sepsis, 85% (171/200) of which matched the culture results based on the detection level. A total of 130 samples were negative according to the Tm mapping method, 98% (128/130) of which were also negative based on the culture method. Meanwhile, 70 samples were positive according to the Tm mapping method, and of the 59 suitable for identification, 100% (59/59) exhibited a "match" or "broad match" with the culture or sequencing results. These findings were obtained within three hours of whole blood collection. The Tm mapping method is therefore useful for identifying infectious diseases requiring prompt treatment.
authors.affiliation: Clinical Laboratory Center, Toyama University Hospital, Toyama 930-0194, Japan.
authors.forename: Hideki
authors.initials: H
authors.lastname: Niimi
authors.affiliation: Clinical Laboratory Center, Toyama University Hospital, Toyama 930-0194, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Ueno
authors.affiliation: Clinical Laboratory Center, Toyama University Hospital, Toyama 930-0194, Japan.
authors.forename: Shirou
authors.initials: S
authors.lastname: Hayashi
authors.affiliation: Kitami Information Technology Co., Ltd., Hokkaido 090-0813, Japan.
authors.forename: Akihito
authors.initials: A
authors.lastname: Abe
authors.affiliation: Kitami Information Technology Co., Ltd., Hokkaido 090-0813, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Tsurue
authors.affiliation: Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Ishikawa 921-8836, Japan.
authors.forename: Masashi
authors.initials: M
authors.lastname: Mori
authors.affiliation: Life Science Center, Hokkaido Mitsui Chemicals, Inc., Hokkaido 073-0138, Japan.
authors.forename: Homare
authors.initials: H
authors.lastname: Tabata
authors.affiliation: Life Science Center, Hokkaido Mitsui Chemicals, Inc., Hokkaido 073-0138, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Minami
authors.affiliation: Department of Internal Medicine, University of Iowa Carver College of Medicine, IA 52242, USA, and Iowa City Veterans Affairs Medical Center, IA 52246, USA.
authors.forename: Michihiko
authors.initials: M
authors.lastname: Goto
authors.affiliation: Nagaresugi Geriatric Hospital, Toyama 939-8032, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Akiyama
authors.affiliation: Department of Clinical Infectious Diseases, Toyama University Hospital, Toyama 930-0194, Japan.
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Yamamoto
authors.affiliation: Department of Obstetrics &Gynecology, Toyama University Hospital, Toyama 930-0194, Japan.
authors.forename: Shigeru
authors.initials: S
authors.lastname: Saito
authors.affiliation: Clinical Laboratory Center, Toyama University Hospital, Toyama 930-0194, Japan.
authors.forename: Isao
authors.initials: I
authors.lastname: Kitajima
mesh_terms: D001419:Bacteria / Q000235:genetics; D001424:Bacterial Infections / Q000175:diagnosis / Q000382:microbiology; D006801:Humans; D008828:Microbiological Techniques; D058889:Molecular Typing / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D015203:Reproducibility of Results; D013048:Specimen Handling / Q000379:methods; D013997:Time Factors; D044366:Transition Temperature; D057188:Workflow