title: Clinical usefulness of multiplex PCR lateral flow in MRSA detection: a novel, rapid genetic testing method.
abstract: Methicillin-resistant Staphylococcus aureus (MRSA) with exogenous cassette DNA containing the methicillin-resistant gene mecA (SCCmec) poses a problem as a drug-resistant bacterium responsible for hospital- and community-acquired infections. The frequency of MRSA detection has recently been increasing rapidly in Japan, and SCCmec has also been classified more diversely into types I-V. A rapid test is essential for early diagnosis and treatment of MRSA infections, but detection by conventional methods requires at least two days. The newly developed multiplex PCR lateral flow method allows specific amplification of femA to detect S. aureus, mecA to detect SCCmec, and kdpC to detect SCCmec type II; moreover, PCR products can be evaluated visually in about 3 h. In the present study, we developed a PCR lateral flow method for MRSA using this method and investigated its clinical usefulness in the detection of MRSA. The results showed a diagnostic concordance rate of 91.7% for MRSA and methicillin-susceptible S. aureus between bacteriological examination and PCR lateral flow, and a high level of specificity in PCR lateral flow. In addition, a higher detection rate for S. aureus using the same sample was observed for PCR lateral flow (70.2%) than for bacteriological tests (48.6%). The above results show that PCR lateral flow for MRSA detection has high sensitivity, specificity, and speed, and its clinical application as a method for early diagnosis of MRSA infections appears to be feasible.
authors: Department of Clinical Laboratory, Kitasato University Hospital, 1-15-1 Kitasato, Minami-ku, Sagamihara-shi, Kanagawa 228-8555, Japan. shin0225@kitasato-u.ac.jp
authors: Shin
authors: S
authors: Nihonyanagi
authors: Yuhsaku
authors: Y
authors: Kanoh
authors: Kiyomi
authors: K
authors: Okada
authors: Toshiki
authors: T
authors: Uozumi
authors: Yukumasa
authors: Y
authors: Kazuyama
authors: Tokiko
authors: T
authors: Yamaguchi
authors: Nobuhiko
authors: N
authors: Nakazaki
authors: Keizou
authors: K
authors: Sakurai
authors: Yasuyoshi
authors: Y
authors: Hirata
authors: Shinichi
authors: S
authors: Munekata
authors: Shinichi
authors: S
authors: Ohtani
authors: Tsuyoshi
authors: T
authors: Takemoto
authors: Yuki
authors: Y
authors: Bandoh
authors: Tohru
authors: T
authors: Akahoshi
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000235:genetics; D018076:DNA, Complementary; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D046915:Penicillin-Binding Proteins; D011494:Protein Kinases / Q000235:genetics; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000235:genetics / Q000382:microbiology*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D018076:DNA, Complementary; C096686:FemA protein, Bacteria; D046915:Penicillin-Binding Proteins; C487789:mecA protein, Staphylococcus aureus; D011494:Protein Kinases; C075616:kdpD protein, Bacteria