ABSTRACT
<p><b>OBJECTIVE</b>To establish a rapid, sensitive and specific real-time PCR method for detection of Human Herpesvirus-6 (HHV-6).</p><p><b>METHODS</b>According to the reference, a pair of primers and a probe were designed located in U65-66 gene and to set up the standards. We established a real-time RT-PCR method for detection of HHV-6, and to verify the specificity, sensitivity, reproducibility.</p><p><b>RESULTS</b>The correlation coefficient was 0.999, E = 97.9%, the coefficient of variation values of Ct were 0.61% and 3.13% in real-time PCR assay for inter and intra assay, respectively. The results of all viruses were negative except of HHV-6 for the assay. The quantitative detection limit of the assay was 3 x 10(0) copies/microl.</p><p><b>CONCLUSION</b>The real-time PCR assay is highly specific, sensitive and reproducible, which can be used to quatitative detecting clinical samples.</p>
Subject(s)
Humans , Herpesvirus 6, Human , Genetics , Real-Time Polymerase Chain Reaction , Methods , Reproducibility of Results , Reverse Transcriptase Polymerase Chain Reaction , MethodsABSTRACT
<p><b>OBJECTIVE</b>To explore the viral etiology of acute low respiratory tract infection (ALRTI) among hospitalized children in Changsha of Hunan Province of China.</p><p><b>METHODS</b>Nasopharyngeal aspirates were collected from 1165 hospitalized children with ALRTI in Changsha from September 2007 to August 2008. Respiratory syncytin virus (RSV), human rhinovirus (HRV), influenza virus A (IFVA), influenza virus B (IFVB), parainfluenza 1-3 (PIV 1-3), human metapneumovirus (hMPV), human coronaviruses NL63 (HCoV-NL63), and human coronaviruses HKU1 (HCoV-HKU1) were detected by reverse transcription polymerase chain reaction (RT-PCR). Adenovirus (ADV) and human bocavirus (HBoV) were detected by standard polymerase chain reaction (PCR). WU polyomaviruses (WUPyV) and KI polyomaviruses(KIPyV) were detected by nested PCR. The positive samples further underwent genetic sequencing.</p><p><b>RESULTS</b>Among the 1165 nasopharyngeal aspirates, viruses were detected in 871 samples (74.76%), among which RSV (27.03%) was the most common virus, followed by HRV (17.33%), PIV3 (13.73%), HBoV (8.67%) and hMPV (6.52%). The overall positive rate of viral detection showed no significant differences between males and females (X2=2.241, P=0.134), whereas the positive rates of PIV3, hMPV, and HBoV in males were higher than in females. The positive rate of viral detection showed significant differences among different age groups (X2=10.934, P=0.027), and the highest positive rate was noted in the age group of 6 months to 1 year. Furthermore, the overall positive rate of viral detection showed a significant difference in term of seasonal distribution, with a peak prevalence in winter.</p><p><b>CONCLUSIONS</b>Virues predominate in the etiology of pediatric ALRTI in Changsha, and RSV, HRV and PIV3 are the main viruses for ALRTI. HBoV and hMPV have become increasingly important. Viral infection-associated ALRTI shows a prevail in the age group of 6 months to 1 year as well as in winter.</p>
Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Age Distribution , Child, Hospitalized , Nasopharynx , Virology , Respiratory Tract Infections , Virology , Seasons , Sex Distribution , VirusesABSTRACT
75%) in P 21 d in group HO.2.The apoptotic index in group HO was higher than that of group Air on P 3 d(Pa