RESUMEN
This article has been retracted: please see Elsevier Policy on Article Withdrawal (https://www.elsevier.com/about/our-business/policies/article-withdrawal). This article has been retracted at the request of the authors. There is significant discrepancy in the genomic characteristics between the currently and previously dominant GBS (2018) and previously dominant GBS (2013-2014). The dramatically rapid and unexpected evolution of GBS strains has led to the significant discrepancy from recent findings which makes all the authors strongly concerned that this will influence the accuracy and validity of GBS treatment and therapy if based on the current manuscript. For example, the genomic difference between the currently prevalent type (II and III) and previously prevalent type (III) is considerably diverse, for which the pathogenic and virulent characteristics of the strains are very different. As all authors have a strong sense of responsibility and expertise in clinical microbiology, agreed by all authors, on behalf of all authors of this manuscript, the authors consequently request for article withdrawal for this manuscript.
Asunto(s)
Genotipo , Serogrupo , Infecciones Estreptocócicas/epidemiología , Infecciones Estreptocócicas/microbiología , Streptococcus agalactiae/genética , Streptococcus agalactiae/aislamiento & purificación , Streptococcus agalactiae/patogenicidad , China/epidemiología , Humanos , Lactante , Recién Nacido , Meningitis/epidemiología , Meningitis/microbiología , Tipificación de Secuencias Multilocus , Reacción en Cadena de la Polimerasa Multiplex , Neumonía/epidemiología , Neumonía/microbiología , Prevalencia , Sepsis/epidemiología , Sepsis/microbiologíaRESUMEN
BACKGROUND: Recognized as a resistance mechanism responsible for the emergence and prevalence of antimicrobial resistance, integron is widely distributed and spread among clinical microorganisms and play a key role in the dissemination of such antimicrobial resistance, which may eventually contribute to the unleashing of "Super Bugs" In this study, detection assays based on loop-mediated isothermal amplification (LAMP) methodologies targeting on class 1 to class 3 integrase genes was developed and evaluated. METHODS: LAMP methodology was employed to develop novel detection assays on class 1, 2 and 3 integrons. Firstly, this protocol was specifically designed to detect such integrons by targeting integrase genes intI1, intI2 and intI3. Development, evaluation and optimization of such LAMP assays was studied, including the reaction temperature, volumn, time, sensitivity and specificity of both primers and targets. A total of 1082 strains, including 397 integron positive and 685 integron negative microorganisms, were included for the application verification of the established LAMP assays. RESULTS: The indispensability of each primer was confirmed, and the optimal amplification was obtained under 63 °C for 45 min, with 25 µl reaction found to be the most cost-efficient volume. As application was concerned, all of the 397 integron-positive isolates yielded positive amplicons and other 685 integron-negative bacteria were negative for the integron-LAMP assays, revealing totaling 100% detection rate and specificity. CONCLUSIONS: The established integron-LAMP assays was demonstrated to be a valid and rapid detection method for integrons screening, which may aid in both the laboratory and clinical integron screening for microorganisms.
Asunto(s)
Farmacorresistencia Bacteriana/genética , Integrones/genética , Secuencias Repetitivas Esparcidas/genética , Técnicas de Amplificación de Ácido Nucleico/métodos , Antibacterianos , Bacterias/genética , ADN Bacteriano/genética , ADN Bacteriano/aislamiento & purificación , Calor , Integrasas/clasificación , Integrasas/genética , Sensibilidad y Especificidad , Virulencia/genéticaRESUMEN
In the Viable but Non-Culturable (VBNC) state, microorganisms may survive under severe external environment. In this study, the specificity and sensitivity of PMA-LAMP assay on the detection of Vibrio Parahemolyticus (V. parahemolyticus) has been developed and evaluated, with further application on a number of food-borne V. parahemolyticus strains. Six primers were designed for recognizing 8 distinct targeting on tlh, tdh and trh gene. Through specific penetration through the damaged cell membrane of dead cells and intercalating into DNA, PMA could prevent DNA amplification of dead bacteria from LAMP, which enabled the differentiation of bacteria between VBNC state and dead state. The established PMA-LAMP showed significant advantage in rapidity, sensitivity and specificity, compared with regular PCR assay. The applicability had also been verified, demonstrating the PMA-LAMP was capable of detection on V. parahaemolyticus.