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.
Eur J Pediatr. 2018 Jan;177(1):85-93. doi: 10.1007/s00431-017-3036-3. Epub 2017 Oct 31.
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.
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.
本研究旨在比较两种用于鉴定与新生儿败血症相关细菌的方法：血液培养和广谱 16S rDNA-DGGE。
从有（n=15）或无（n=7）败血症症状和体征的新生儿中获取 22 份血样。使用两种不同的方法对血液样本进行筛选以鉴定致病菌：（1）细菌培养和（2）16S rDNA-DGGE 的可变 V3 区扩增。
血液培养分析的阳性率为 40%，而 16S rDNA-DGGE 的阳性率为 87%。所有 16S rDNA-DGGE 阳性样本均与新生儿败血症的其他一些迹象有关。
我们的研究表明，16S rDNA-DGGE 的分子方法比细菌培养法鉴定出两倍多的致病菌，包括与新生儿细菌性败血症发展相关的复杂细菌群落。
•新生儿败血症影响 NICU 中 2.3%的出生，具有高死亡风险。•有证据支持使用分子方法替代血液培养来鉴定与新生儿败血症相关的细菌。
•DGGE 凝胶是鉴定新生儿血液样本中细菌的一种很好的方法学方法。•本研究描述了 DGGE 凝胶获得的电泳迁移模式，并能够确定与新生儿败血症发展相关的细菌类型。
