Tracheal aspirate as a substrate for polymerase chain reaction detection of viral genome in childhood pneumonia and myocarditis.
BACKGROUND
Infectious respiratory disorders are important causes of childhood morbidity and mortality. Viral causes are common and may lead to rapid deterioration, requiring mechanical ventilation; myocardial dysfunction may accompany respiratory decompensation. The etiologic viral diagnosis may be difficult with classic methods. The purpose of this study was to evaluate polymerase chain reaction (PCR) as a diagnostic method for identification of causative agents.


RESULTS
PCR was used to amplify sequences of viruses known to cause childhood viral pneumonia and myocarditis. Oligonucleotide primers were designed to amplify specific sequences of DNA virus (adenovirus, cytomegalovirus, herpes simplex virus, and Epstein-Barr virus) and RNA virus (enterovirus, respiratory syncytial virus, influenza A, and influenza B) genomes. Tracheal aspirate samples were obtained from 32 intubated patients and nucleic acid extracted before PCR. PCR results were compared with results of culture, serology, and antigen detection methods when available. In cases of myocarditis (n=7), endomyocardial biopsy samples were analyzed by PCR and compared with tracheal aspirate studies. PCR amplification of viral genome occurred in 18 of 32 samples (56%), with 3 samples PCR positive for 2 viral genomes. Amplified viral sequences included RSV (n=3), enterovirus (n=5), cytomegalovirus (n=4), adenovirus (n=3), herpes simplex virus (n=2), Epstein-Barr virus (n=1), influenza A (n=2), and influenza B (n=1). All 7 cases of myocarditis amplified the same viral genome from heart as found by tracheal aspirate.


CONCLUSIONS
PCR is a rapid and sensitive diagnostic tool in cases of viral pneumonia with or without myocarditis, and tracheal aspirate appears to be excellent for analysis.
Akhtar
N
N
Department of Pediatrics, Section of Critical Care, Baylor College of Medicine, Houston, TX 77030, USA.
Ni
J
J
Stromberg
D
D
Rosenthal
G L
GL
Bowles
N E
NE
Towbin
J A
JA
D000257:Adenoviridae Infections / Q000175:diagnosis / Q000821:virology*; D000260:Adenoviruses, Human / Q000235:genetics / Q000302:isolation & purification; D000293:Adolescent; D001483:Base Sequence; D001706:Biopsy; D001826:Body Fluids / Q000821:virology*; D002648:Child; D002675:Child, Preschool; D003587:Cytomegalovirus / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000302:isolation & purification*; D004699:Endocardium / Q000821:virology; D004770:Enterovirus / Q000235:genetics / Q000302:isolation & purification; D004769:Enterovirus Infections / Q000175:diagnosis / Q000821:virology*; D004797:Enzyme-Linked Immunosorbent Assay; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000821:virology*; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D009980:Influenza A virus / Q000235:genetics / Q000302:isolation & purification; D009981:Influenza B virus / Q000235:genetics / Q000302:isolation & purification; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology*; D008969:Molecular Sequence Data; D009205:Myocarditis / Q000175:diagnosis / Q000821:virology*; D011024:Pneumonia, Viral / Q000175:diagnosis / Q000821:virology*; D016133:Polymerase Chain Reaction; D012367:RNA, Viral / Q000302:isolation & purification*; D012127:Respiratory Distress Syndrome, Newborn / Q000821:virology; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis / Q000821:virology*; D012136:Respiratory Syncytial Viruses / Q000235:genetics / Q000302:isolation & purification; D018139:Simplexvirus / Q000235:genetics / Q000302:isolation & purification; D013396:Suction; D014132:Trachea / Q000821:virology*; D014776:Virus Cultivation
D004279:DNA, Viral; D012367:RNA, Viral