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. Objectives: 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 × 10⁶ 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.

Prevalence of Human Herpes Viruses in Bronchoalveolar Lavage of Critically Ill Children Undergoing Mechanical Ventilation at a Pediatric Intensive Care Unit

“…Diagnostic virology has become one of the integral parts of managing patients with respiratory infection (1)(2)(3)(4)(5). Along with recent advantages in diagnosis (6), new challenges have occurred in the management of viral infections (true disease versus colonization), especially among high risk patients (such as immunocompromised hosts).…”

“…Human herpes viruses (HHVs) are one of the most common causes of respiratory viral infections in immunocompromised patients (1,20). Although cytomegalovirus (CMV) is a known cause (21,22), other members of this family may cause pneumonia in both healthy and immunocompromised individuals (3,23,24).…”

“…Although cytomegalovirus (CMV) is a known cause (21,22), other members of this family may cause pneumonia in both healthy and immunocompromised individuals (3,23,24). Patients with acute respiratory distress syndrome (ARDS) and acute lung injury (ALI) or viral pneumonia of unknown etiology should be examined for HHVs (1,24).…”

Viral Pneumonia in Severe Respiratory Failure

“…In this issue of Respiration , Tachikawa et al [16] report the use of rapid screening qualitative PCR for identifying herpes viral DNA in respiratory samples from 87 patients with acute lung injury or ARDS, followed by quantitative PCR of the viral load from the samples where Herpesviridae was detected. This was a mixed group of patients with respiratory failure, approximately 56% of whom were immunocompromised due to chronic steroid use or acquired immunodeficiency; 31% were intubated.…”

“…Third, none of the bronchoalveolar fluid levels correlated with blood markers of viremia, reiterating the importance of site-specific sampling. Finally, Tachikawa et al provide a putative diagnostic threshold viral load for CMV in bronchoalveolar fluid with 87.5% sensitivity and 84.6% specificity, for likely or probable viral pneumonia [16]. …”