+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Molecular typing of adenoviruses isolated from clinical specimens by PCR and DNA sequencing methods



Molecular typing of adenoviruses isolated from clinical specimens by PCR and DNA sequencing methods



Mikrobiyoloji Bulteni 46(4): 607-613



Adenoviruses are responsible for a broad spectrum of diseases, including upper and lower respiratory tract infections (URTIs and LRTIs, respectively), conjunctivitis, gastroenteritis, and hemorrhagic cystitis. The aim of this study was to determine the adenovirus (AdV) types isolated from clinical specimens by polymerase chain reaction (PCR) and DNA sequencing methods. A total of 22 AdV strains isolated between January 1st 2011 to May 31th 2011, from various samples (295 nasopharyngeal swabs, 42 conjunctival swabs, 13 stool) sent to our routine virology laboratory were included in the study. Of the 22 patients whose samples yielded adenovirus positivity, 8 were adult (4 were male; median age: 32.5 years) and 14 (7 were male; median age: 1 year) were children. Those specimens (14 nasopharyngeal swabs, 7 conjunctival swabs, 1 stool) were obtained from patients with URTIs (n= 6), LRTIs (n= 8), conjunctivitis (n= 7) and gastroenteritis (n= 1). For the isolation and identification of adenoviruses, rapid (shell vial) cell culture and direct immunofluorescence antibody methods were used, respectively. Molecular typing of adenoviruses were performed by PCR and sequencing of a partial region (hipervariable region 1-6) of the hexon gene. PCR primers (Adhex F1, Adhex R1) used for DNA amplification were from those described by Lu and Erdman, previously. If insufficient DNA was amplified from the first reaction for sequencing, a nested PCR was performed using Adhex F2 and Adhex R2 primers. Sequencing was performed using the amplification primers and Sequence Reagent Mix-DYEnamic ET Terminator Cycle Sequencing Kit (Amersham Pharmacia Biotech Inc, USA) on ABI PRISM 310 Genetic Analyzer (Applied Biosystems, USA). Obtained adenovirus sequences were typed by BLAST analysis and three AdV types namely type 3, 4, and 8 were identified. In our study, AdV type 3 was detected in a gastroenteritis case and six cases with URTIs and LRTIs (n= 7, 31.8%). AdV type 8 was identified as the cause of conjunctivitis in seven patients and of URTIs and LRTIs in five patients (n= 12, 54.5%). AdV type 4 was found to be associated with URTI in one, and LRTIs in two patients (n= 3; 13.7%). Our data indicated that AdV type 8 was the most prevalent type in patients with conjunctivitis and URTIs, while AdV type 3 was the most prevalent type in patients with LRTI. BLAST analysis was thought to be useful for the molecular typing of adenoviruses. In conclusion, advanced studies with large number of specimens are necessary to achive a reliable, detailed national adenovirus database.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 054451088

Download citation: RISBibTeXText

PMID: 23188574


Related references

Molecular identification and typing of enteroviruses isolated from clinical specimens. Journal of Clinical Microbiology 40(12): 4554-4560, 2002

Molecular typing of human adenoviruses by PCR and sequencing of a partial region of the hexon gene. Archives of Virology 151(8): 1587-1602, 2006

Molecular typing of Vibrio vulnificus strains isolated from clinical specimens by pulsed-field gel electrophoresis and random amplified polymorphic DNA. Abstracts of the Interscience Conference on Antimicrobial Agents & Chemotherapy 37: 348, 1997

Molecular typing of vibrio vulnificus isolated from clinical specimens by pulsed-field gel electrophoresis and random amplified polymorphic DNA analysis. Japanese Journal of Infectious Diseases 52(2): 38-41, 1999

Molecular typing of fowl adenoviruses, isolated in Hungary recently, reveals high diversity. Veterinary Microbiology 167(3-4): 357-363, 2013

Genetic characterization of Methicillin-Resistant Staphylococcus aureus (MRSA) isolated from clinical specimens of various animal species investigated by molecular typing procedures. 2007

Comparison of the ELVIS typing test with traditional methods for detection and typing of herpes simplex virus from clinical specimens. Abstracts of the Interscience Conference on Antimicrobial Agents & Chemotherapy 37: 230, 1997

Genomic typing of BK virus in clinical specimens by direct sequencing of polymerase chain reaction products. Journal Of Medical Virology. 41(1): 11-17, 1993

Detection and typing of human herpesvirus 6 by molecular methods in specimens from patients diagnosed with encephalitis or meningitis. Journal of Clinical Microbiology 45(12): 3972-3978, 2007

Parechovirus typing in clinical specimens by nested or semi-nested PCR coupled with sequencing. Journal of Clinical Virology 48(3): 202-207, 2010

Typing of coagulase negative staphylococci isolated from clinical specimens and detection of slime production. Mikrobiyoloji Bulteni 30(1): 19-24, 1996

Typing of human adenoviruses in specimens from immunosuppressed patients by PCR-fragment length analysis and real-time quantitative PCR. Journal of Clinical Microbiology 44(8): 2808-2815, 2006

Molecular detection and typing of duck hepatitis A virus directly from clinical specimens. Veterinary Microbiology 131(3-4): 247-257, 2008

Study on serotyping, phage typing and drug resistance of Serratia marcescens isolated from clinical specimens. Nihon Hinyokika Gakkai Zasshi. Japanese Journal of Urology 71(11): 1313-1327, 1980