Detection and differentiation of human herpesviruses 1-5 by consensus primer PCR and RFLP.

Eight human viruses of the Herpesviridae family represent a significant public health problem world-wide. Detection and typing of five of the human herpesviruses (HSV-1, HSV-2, VZV, EBV, and CMV) was performed by applying a consensus primer polymerase chain reaction (PCR). The amplified PCR products from the five human herpesviruses were typed based on their restriction enzyme digestion polymorphism with Hinf I and Alu I. Fifteen clinically suspected specimens from herpesvirus-infected patients were also evaluated. A fragment of the DNA polymerase gene from each of the five human herpesviruses was successfully amplified by the set of consensus primers. Their amplicons obtained by PCR from the template DNAs were subjected to restriction endonuclease digestion and human herpesviruses 1-5 could be clearly differentiated and typed. This method can be used to detect and differentiate between the five human herpesviruses in clinical specimens. This study demonstrates the value of testing for five human herpesviruses by consensus PCR and restricted fragment length polymorphism (RFLP). These procedures are simple and straightforward techniques for the investigation of clinical specimens.

Plasma endothelin-1 in infants and young children with acute bronchiolitis and viral pneumonia.

Respiratory syncytial virus (RSV) Infections that occur during the first three years of life have been demonstrated to be associated with the development of childhood asthma. The mechanism of virus-triggered airway inflammation Is not fully understood. Endothelin-1 is a potent bronchoconstrictor involved in many diseases including respiratory tract infections. Infants and young children diagnosed with either viral pneumonia or acute bronchiolitis, their age ranging between 2 months and 3 years, were recruited into this study. Nasopharyngeal aspirates were taken for detection of respiratory virus by antigen immunofluorescence stain, RT-PCR analysis and viral culture. Plasma endothelin-1 (ET-1) was measured by using a commercially available enzyme-linked immunosorbent assay (ELISA). Ten of the nineteen infants and children (52%) were positive for RSV infection, one co-infected with influenza A. Nine Infants (90%) were positive for RSV subtype A. There was only one infant with subtype B. One of the RSV negative individuals was positive for influenza A. In addition, we recruited 10 patients without chronic underlying or respiratory tract illness as controls. ET-1 levels were significantly increased in RSV infection compared to the controls (3.6 +/- 1.2 and 1.2 +/- 1 pg/ml, respectively (p < 0.05). In conclusion, infants and young children who are infected with RSV have an increase in circulating plasma endothelin-1. This in turn may contribute to the subsequent development of childhood asthma.