2012, Number 3
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Rev Invest Clin 2012; 64 (3)
The usefulness of the buffy coat smear and panbacterial polymerase chain reaction in early diagnosis of neonatal sepsis
García-Elorriaga G, Cortés-Torres N, Ballesteros-del-Olmo JC, Del Rey-Pineda G, González-Bonilla C
Language: English
References: 27
Page: 275-283
PDF size: 183.09 Kb.
ABSTRACT
Objective. In this study are evaluated the usefulness of the
buffy coat smear and panbacterial polymerase chain reaction
(PCR) as diagnostic tests in the early detection of neonatal
sepsis.
Material and methods. It was studied 49 patients
aged up to 28 days who were hospitalized in the Intensive
Care Unit (ICUs) of the Neonatology, with a clinical diagnosis
of neonatal sepsis and 49 umbilical cord samples of healthy
newborns. Blood cultures and 50 μL of plasma were taken for
the DNA and performance of the broad-range PCR primer
system (panbacterial PCR). Simultaneously, were taken three
capillaries with blood for the leukocyte layer (buffy coat)
smear, we performed three stains: Gram; Löeffler blue methylene
(LBM), and acridine orange (AO). Statistical analysis
included sensitivity, specificity, positive predictive value
(PPV), and negative predictive value (NPV) against the clinical
diagnosis.
Results. With respect to stains of
buffy coat
smear, they resulted very specific, from 90-97%, with 64-75%
sensitivity, 87-94% PPV, and 77-82% NPV. In inverse fashion,
PCR resulted very sensitive at 96%, with 91% specificity,
92% PPV, and 96% NPV.
Conclusions. Buffy coat smear
stains are easy, fast, and specific, while that of PCR was highly
sensitive. Thus, both can be utilized as diagnostic tests.
REFERENCES
Goldstein B, Giroir B, Randolph A. International pediatric sepsis consensus conference: definitions for sepsis and organ dysfunction in pediatrics. Pediatric Crit Care Med 2005; 6: 1-8.
Ballesteros-del-Olmo JC, Merás-Montoya AI. Infecciones en Neonatología en un tercer nivel de atención. Boletín de la Academia Mexicana de Pediatría 2006. Disponible en: http:// www.academiamexicanadepediatria.com.mx/publicaciones/academicos opinan/otras.php
Rodríguez-Weber MA, López-Candiani C, Arredondo-García JL, Gutiérrez-Castrellón P, Sánchez-Arriaga F. Morbilidad y mortalidad por sepsis neonatal en un hospital de tercer nivel de atención. Sal Pub Mex 2003; 45: 90-5.
Sundaram V, Kumar P, Dutta S, Mukhopadhyay K, Ray P, Gautam V, et al. Blood culture confirmed bacterial sepsis in neonates in a North Indian tertiary care center: changes over the last decade. Jpn J Infect Dis 2009; 60: 46-50.
Ballesteros JC, Rodríguez C, Morales M, García-E G, García- RP, Vega-G J, et al. Indicadores de infección temprana en septicemia neonatal. Rev Mex Pediatr 1996; 63: 17-24.
Kumarcs V, Neelagaud YF. Incubation period for culture positivity to detect septicemia in neonates. Indian J Med Microbiol 2005; 23: 270-1.
McCabe W, LaPorte J. Intracellular bacteria in the peripheral blood in Staphylococcal bacteremia. Ann Intern Med 1962; 57: 141-3.
Faden H. Early diagnosis of neonatal bacteremia by buffy coat examination. J Pediatr 1976; 88: 1032-4.
Lemus-Varela ML, Alberto VS, Arriaga-Dávila JJ. Clinical and laboratory parameters in neonatal nosocomial sepsis. Gac Med Mex 2008; 144: 409-11.
Gérard HC, Wang Z, Wang GF, El-Gabalawy H, Goldbach- Mansky R, Li Y, et al. Chromosomal DNA from a variety of bacterial species is present in synovial tissue from patients with various forms of arthritis. Arthritis Rheum 2001; 44: 1689-97.
Dutta S, Narang A, Chakraborty A, Ray P. Diagnosis of neonatal sepsis using universal primer polymerase chain reaction before and after starting antibiotic therapy. Arch Pediatr Adolesc Med 2009; 163: 6-11.
Ohlin A, Bäckman A, Björkqvist M, Mölling P, Jurstrand M, Schollin J. Real-time PCR of the 16S-rRNA gene in the diagnosis of neonatal bacteraemia. Acta Paediatr 2008; 97: 1376-80.
Chomczynski P. A reagent for the single-step simultaneous isolation of RNA, DNA and proteins from cell and tissue samples. Biotechniques 1993; 15: 532-7.
Gutell RR, Larsen N, Woese CR. Lesson from an evolving rRNA: 16S and 23S structures from a comparative perspective. Microbiol Rev 1994; 58: 10-26.
Ponce-de-León S. Como leer revistas médicas para aprender sobre una prueba diagnóstica. Rev Invest Clin 1988; 40: 73-83.
Gutiérrez-Muñoz UH, Gutiérrez-Muñoz J, Rosas-Barrientos V. Factores de riesgo en sepsis neonatal en un hospital de tercer nivel en la Ciudad de México. Revista de Especialidades Médico- Quirúrgicas, ISSSTE 2005; 10: 21-4.
Murgía M, Mancilla J. Programa de actualización continúa en neonatología. Libro 7. México: Intersistemas Editores; 2004.
Manroe BL, Weinberg AG, Rosenfeld CR, Browne R. The neonatal blood count in health and disease. I. Reference values for neutrophilic cells. J Pediatr 1979; 95: 89-98.
Isaacman DJ, Karasic RB, Reynolds EA, Krost SI. Effect of number of blood cultures and volume of blood and detection of bacteremia in children. J Pediatr 1996; 128: 190-5.
Mangurten HH, Le Beau LJ. Diagnosis of neonatal bacteremia by a microblood culture technique. J Pediatr 1997; 90: 990.
Domínguez-Sosa J, Vila-Ruiz F, Setién-Castillo IA. Prevalencia y resistencia bacteriana en una unidad de cuidados intensivos neonatales. Enfermedades Infecciosas y Microbiología 2005; 25(3).
Ballesteros-del-Olmo JC, Tena-Reyes D, García-Elorriaga G, Vega-García A, Ramírez-Ortiz M, Sosa-Maldonado J, et al. Sensibilidad y especificidad del frotis de buffy coat teñido con Gram en el diagnóstico de sepsis neonatal. Rev Mex Pediatr 2008; 75: 97-102.
Zamora-Castorena S, Murgía-de Sierra MF. Cinco años de experiencia con sepsis neonatal en un centro pediátrico. Rev Invest Clin 1998; 50: 463-70.
Kostiala A, Jormalainen S, Kosunen T. Detection of experimental bacteremia and fungemia by examination of buffy coat prepared by a micromethod. Am J Clin Pathol 1979; 72: 437-43.
Melly MA, Thomison JB, Rogers DE. Fate of staphylococci within human leukocytes. J Exp Med 1960; 112: 1121-30.
Figueras-Aloy J, Gómez-López L, Rodríguez-Miguélez JM, Salvia-Roiges MD, Jordán-García I, Ferrer-Codina I, et al. Serum soluble ICAM-1, VCAM-1, L-selectin, and P-selectin levels as markers of infection and their relation to clinical severity in neonatal sepsis. Am J Perinatol 2007; 24: 331-8.
Barman P, Sengupta S, Singh S. Study of a novel method to assist in early reporting of sepsis from the microbiology laboratory. J Infect Dev Ctries 2010; 4(12): 822-7.