BACKGROUND: Conventional blood culture is still the gold standard for sepsis diagnosis but results are not immediately available and pathogens are only detected in approximately 25% of cases. New molecular assays for the detection of blood stream pathogens are promising diagnostic tools. OBJECTIVES: The aim of the study was to adapt and evaluate a multiplex PCR system using 100 µl blood. - METHODS: 46 blood specimens of very low birth weight infants (818 ± 242 g) with suspected sepsis were analyzed using the Roche SeptiFast MGRADE PCR with a modified DNA extraction protocol and software handling tool for decreased blood volume requirements. RESULTS: In the non-infected group, 5/21 infants had a positive PCR result with coagulase-negative staphylococci. All pathogens detected in the blood culture positive group (n = 15) were also detected by PCR. In addition, 4/6 patients had a positive PCR result in the clinical sepsis group (clinical and laboratory signs of sepsis but negative blood culture). Overall, the PCR was demonstrated to have a higher sensitivity (90.5%; 95%CI 68.2-98.3%) in comparison to blood culture (71.4%; 95%CI 47.7-87.8%) including clinical sepsis cases, even though it had a lower specificity (80.0%; 95%CI 58.7-92.4% versus 100.0%; 95%CI 83.4-100.0%). CONCLUSIONS: These first data demonstrate the usability and potential benefit of this multiplex PCR using a modified DNA extraction for the rapid detection of nosocomial sepsis in preterm infants in addition to blood culture.
BACKGROUND: Conventional blood culture is still the gold standard for sepsis diagnosis but results are not immediately available and pathogens are only detected in approximately 25% of cases. New molecular assays for the detection of blood stream pathogens are promising diagnostic tools. OBJECTIVES: The aim of the study was to adapt and evaluate a multiplex PCR system using 100 µl blood. - METHODS: 46 blood specimens of very low birth weight infants (818 ± 242 g) with suspected sepsis were analyzed using the Roche SeptiFast MGRADE PCR with a modified DNA extraction protocol and software handling tool for decreased blood volume requirements. RESULTS: In the non-infected group, 5/21 infants had a positive PCR result with coagulase-negative staphylococci. All pathogens detected in the blood culture positive group (n = 15) were also detected by PCR. In addition, 4/6 patients had a positive PCR result in the clinical sepsis group (clinical and laboratory signs of sepsis but negative blood culture). Overall, the PCR was demonstrated to have a higher sensitivity (90.5%; 95%CI 68.2-98.3%) in comparison to blood culture (71.4%; 95%CI 47.7-87.8%) including clinical sepsis cases, even though it had a lower specificity (80.0%; 95%CI 58.7-92.4% versus 100.0%; 95%CI 83.4-100.0%). CONCLUSIONS: These first data demonstrate the usability and potential benefit of this multiplex PCR using a modified DNA extraction for the rapid detection of nosocomial sepsis in preterm infants in addition to blood culture.
Authors: Kristina A Tendl; Stefan M F Schulz; Thomas P Mechtler; Adele Bohn; Thomas Metz; Susanne Greber-Platzer; David C Kasper; Kurt R Herkner; Chike B Item Journal: Epigenetics Date: 2013-10-17 Impact factor: 4.528
Authors: Javier Ortiz Ibarra; Pablo Trevino Valdez; Ema Valenzuela Mendez; Ana Limon Rojas; Gabriel Lara Flores; Adrian Ceballos Bocanegra; Iyari Morales Mendez; Luis Fernandez Carrocera; Emilia Covian Molina; Jesus Reyna Figueroa Journal: Iran J Pediatr Date: 2015-01-17 Impact factor: 0.364