title: Urine Nested Polymerase Chain Reaction in Neonatal Septicemia.
abstract: This cross-sectional study was done to evaluate diagnostic efficacy of urine nested polymerase chain reaction (PCR) using broad-range 16SrDNA PCR-based amplification, followed by restriction analysis and sequencing in neonatal septicemia. The study included 50 babies; 48% had vaginal delivery, 46% were preterm, 20% had a history of prolonged rupture of membranes and 56% were low birth weight (≤2500 g). Clinical presentations were lethargy (96%), respiratory distress (80%) and bleeding diathesis (16%). Absolute neutrophil count <1800/mm(3) was observed in 60%, and positive C-reactive protein in 46%. Thirty neonates had positive blood culture, and Klebsiella pneumoniae (22%) was the predominant organism. Nested urine PCR was positive in 38 (76%) and detected bacterial DNA in 8 neonates with negative blood cultures. The sensitivity, specificity, positive predictive value, negative predictive value and accuracy of nested PCR were 100, 60, 78.9, 100 and 84%, respectively, compared with blood culture. Nested PCR can detect most bacteria in single assay and identify unusual and unexpected causal agents.
authors.affiliation: Department of Paediatrics, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.
authors.forename: B K
authors.initials: BK
authors.lastname: Das
authors.affiliation: Department of Paediatrics, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Shipra
authors.initials: S
authors.lastname: Suri
authors.affiliation: Department of Microbiology, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India.
authors.forename: Gopal
authors.initials: G
authors.lastname: Nath
authors.affiliation: Department of Paediatrics, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221005, India rajnitip@gmail.com.
authors.forename: Rajniti
authors.initials: R
authors.lastname: Prasad
mesh_terms: D002097:C-Reactive Protein / Q000032:analysis*; D003430:Cross-Sectional Studies; D004247:DNA / Q000032:analysis; D004269:DNA, Bacterial; D005260:Female; D020459:Genes, rRNA / Q000235:genetics; D006801:Humans; D007231:Infant, Newborn; D007711:Klebsiella pneumoniae / Q000302:isolation & purification; D007958:Leukocyte Count; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012336:RNA, Ribosomal, 16S / Q000032:analysis*; D012680:Sensitivity and Specificity; D018805:Sepsis / Q000175:diagnosis* / Q000382:microbiology / Q000652:urine*; D017422:Sequence Analysis, DNA; D013211:Staphylococcus aureus / Q000302:isolation & purification; D014556:Urine / Q000382:microbiology*
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S; D002097:C-Reactive Protein; D004247:DNA
keywords: CRP (C-reactive protein); NPV (negative predictive value); PPV (positive predictive value); Septicemia; neonates; polymerase chain reaction (PCR)