title: Detection of herpes viruses by multiplex and real-time polymerase chain reaction in bronchoalveolar lavage fluid of patients with acute lung injury or acute respiratory distress syndrome.
abstract: BACKGROUND
  Human herpes viruses (HHVs) are important pathogens in acute lung injury (ALI) and acute respiratory distress syndrome (ARDS). Rapid and efficient diagnostic tools are needed to detect HHVs in the lung in ALI/ARDS patients.
  
  
  OBJECTIVE
  This study aimed to evaluate the usefulness of multiplex and real-time polymerase chain reaction (PCR) analysis of bronchoalveolar lavage fluid (BALF) for detecting HHV reactivation in ALI/ARDS patients.
  
  
  METHODS
  Between August 2008 and July 2012, eighty-seven BALF samples were obtained from ALI/ARDS patients with unknown etiology and analyzed for HHVs. The types of HHVs in the BALF samples were determined using qualitative multiplex PCR followed by quantitative real-time PCR.
  
  
  RESULTS
  Multiplex PCR identified herpes simplex virus type 1 (HSV-1) (n = 11), Epstein-Barr virus (EBV) (n = 16), cytomegalovirus (CMV) (n = 21), HHV type 6 (HHV-6) (n = 2), and HHV-7 (n = 1) genomic DNA in 35 (40%) of the BALF samples, including 14 (16%) samples containing 2 or 3 HHV types. CMV and EBV reactivation was rare in immunocompetent patients, whereas reactivation of HSV-1 was predominantly observed in intubated patients regardless of their immune status. Overall, HHVs were almost exclusively found in patients with immunosuppression or endotracheal intubation. Real-time PCR detected 0.95-1.59 × 10(6) copies of viral DNA/μg human genome DNA, and HSV-1 (n = 4), CMV (n = 9), and HHV-6 (n = 1) were identified as potentially pathogenic agents.
  
  
  CONCLUSIONS
  The implementation of multiplex and real-time PCR of BALF was feasible in ALI/ARDS patients, which allowed efficient detection and quantification of HHV DNA.
authors: Department of Respiratory Medicine, Kobe City Medical Center General Hospital, Kobe, Japan.
authors: Ryo
authors: R
authors: Tachikawa
authors: Keisuke
authors: K
authors: Tomii
authors: Ryutaro
authors: R
authors: Seo
authors: Kazuma
authors: K
authors: Nagata
authors: Kyoko
authors: K
authors: Otsuka
authors: Atsushi
authors: A
authors: Nakagawa
authors: Kojiro
authors: K
authors: Otsuka
authors: Hisako
authors: H
authors: Hashimoto
authors: Ken
authors: K
authors: Watanabe
authors: Norio
authors: N
authors: Shimizu
mesh_terms: D055371:Acute Lung Injury / Q000821:virology*; D000368:Aged; D001992:Bronchoalveolar Lavage Fluid / Q000821:virology*; D005260:Female; D006564:Herpesviridae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D060885:Multiplex Polymerase Chain Reaction; D011024:Pneumonia, Viral / Q000175:diagnosis*; D060888:Real-Time Polymerase Chain Reaction; D012128:Respiratory Distress Syndrome / Q000821:virology*; D012189:Retrospective Studies