Multiplex MassTag-PCR for respiratory pathogens in pediatric nasopharyngeal washes negative by conventional diagnostic testing shows a high prevalence of viruses belonging to a newly recognized rhinovirus clade.
BACKGROUND
Respiratory infections are the most common infectious diseases in humans worldwide and are a leading cause of death in children less than 5 years of age.


OBJECTIVE
Identify candidate pathogens in pediatric patients with unexplained respiratory disease.


METHODS
Forty-four nasopharyngeal washes collected during the 2004-2005 winter season from pediatric patients with respiratory illnesses that tested negative for 7 common respiratory pathogens by culture and direct immunofluorescence assays were analyzed by MassTag-PCR. To distinguish human enteroviruses (HEV) and rhinoviruses (HRV), samples positive for picornaviruses were further characterized by sequence analysis.


RESULTS
Candidate pathogens were detected by MassTag PCR in 27 of the 44 (61%) specimens that previously were rated negative. Sixteen of these 27 specimens (59%) contained picornaviruses; of these 9 (57%) contained RNA of a recently discovered clade of rhinoviruses. Bocaviruses were detected in three patients by RT-PCR.


CONCLUSIONS
Our study confirms that multiplex MassTag-PCR enhances the detection of pathogens in clinical specimens, and shows that previously unrecognized rhinoviruses, that potentially form a species HRV-C, may cause a significant amount of pediatric respiratory disease.
Dominguez
Samuel R
SR
Department of Pediatrics, The Children's Hospital, University of Colorado Denver School of Medicine, Aurora, CO, USA.
Briese
Thomas
T
Palacios
Gustavo
G
Hui
Jeffrey
J
Villari
Joseph
J
Kapoor
Vishal
V
Tokarz
Rafal
R
Glodé
Mary P
MP
Anderson
Marsha S
MS
Robinson
Christine C
CC
Holmes
Kathryn V
KV
Lipkin
W Ian
WI
D000208:Acute Disease; D002675:Child, Preschool; D004770:Enterovirus / Q000302:isolation & purification; D004769:Enterovirus Infections / Q000453:epidemiology / Q000821:virology; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D009305:Nasopharynx / Q000821:virology*; D010850:Picornaviridae Infections / Q000453:epidemiology / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D015995:Prevalence; D012141:Respiratory Tract Infections / Q000453:epidemiology / Q000821:virology; D012229:Rhinovirus / Q000145:classification / Q000302:isolation & purification; D014777:Virus Diseases / Q000453:epidemiology / Q000821:virology