Microbiological comparison of blood culture and amplification of 16S rDNA methods in combination with DGGE for detection of neonatal sepsis in blood samples.

Autor: García-Gudiño I; Departamento de Neonatología, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico.; Unidad de Cuidados Intensivos Neonatales, Hospital General Regional 180 Instituto Mexicano del Seguro Social, Tlajomulco de Zúñiga, Jalisco, Mexico., Yllescas-Medrano E; Unidad de Cuidados Intensivos Neonatales, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico., Maida-Claros R; Departamento de Neonatología, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico., Soriano-Becerril D; Departamento de Infectología e Inmunología, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico., Díaz NF; Departamento de Fisiología y Desarrollo Celular, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico., García-López G; Departamento de Fisiología y Desarrollo Celular, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico., Molina-Hernández A; Departamento de Fisiología y Desarrollo Celular, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Mexico City, Mexico., Flores-Herrera O; Departamento de Bioquímica, Facultad de Medicina, UNAM, Mexico City, Mexico., Zavala-Díaz de la Serna FJ; Laboratorio de Biotecnología, Facultad de Ciencias Químicas, Universida Autónoma de Chihuahua, Chihuahua, Mexico., Del Rosario Peralta-Pérez M; Laboratorio de Biotecnología, Facultad de Ciencias Químicas, Universida Autónoma de Chihuahua, Chihuahua, Mexico., Flores-Herrera H; Departmento de Inmunobioquímica, Instituto Nacional de Perinatología 'Isidro Espinosa de los Reyes' INPer, Montes Urales #800 Col. Lomas de Virreyes cp 11000. Tercer piso de la Torre de Investigación, Mexico City, Mexico. h.flores@inper.gob.mx.
Jazyk: angličtina
Zdroj: European journal of pediatrics [Eur J Pediatr] 2018 Jan; Vol. 177 (1), pp. 85-93. Date of Electronic Publication: 2017 Oct 31.
DOI: 10.1007/s00431-017-3036-3
Abstrakt: It is estimated that 15% of all newborns admitted to the neonatal intensive care unit (NICU) for suspected sepsis receive multiple broad-spectrum antibiotics without pathogen identification. The gold standard for bacterial sepsis detection is blood culture, but the sensitivity of this method is very low. Recently, amplification and analysis of the 16S ribosomal DNA (rDNA) bacterial gene in combination with denaturing gradient gel electrophoresis (DGGE) has proven to be a useful approach for identifying bacteria that are difficult to isolate by standard culture methods. The main goal of this study was to compare two methods used to identify bacteria associated with neonatal sepsis: blood culture and broad range 16S rDNA-DGGE. Twenty-two blood samples were obtained from newborns with (n = 15) or without (n = 7) signs and symptoms of sepsis. Blood samples were screened to identify pathogenic bacteria with two different methods: (1) bacteriological culture and (2) amplification of the variable V3 region of 16S rDNA-DGGE. Blood culture analysis was positive in 40%, whereas 16S rDNA-DGGE was positive in 87% of neonatal sepsis cases. All 16S rDNA-DGGE positive samples were associated with some other signs of neonatal sepsis.
Conclusion: Our study shows that the molecular approach with 16S rDNA-DGGE identifies twofold more pathogenic bacteria than bacteriological culture, including complex bacterial communities associated with the development of bacterial sepsis in neonates. What is Known: • Neonatal sepsis affects 2.3% of birth in the NICU with a high mortality risk. • Evidence supports the use of molecular methods as an alternative to blood culture for identification of bacterial associated neonatal sepsis. What is New: • The DGGE gel is a good methodological approach for the identification of bacterial in neonatal blood samples. • This study describes the pattern of electrophoretic mobility obtained by DGGE gels and allows to determine the type of bacteria associated in the development of neonatal sepsis.
Databáze: MEDLINE
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