ABSTRACT
<p><b>OBJECTIVE</b>To establish the real-time fluorescent PCR method for detecting enterovirus, enterovirus 71 and Coxsackievirus A 16 nucleic acid.</p><p><b>METHODS</b>Primers and MGB probe were chosen for virus gene. The samples of 38 HFMD patients were analyzed by TaqMan-MGB PCR technique on a fluorescence real-time PCR instrument,and the results were compared with those by conventional RT-PCR.</p><p><b>RESULTS</b>The real-time fluorescent PCR positive rates of EV, EV71 and Cox A16 were 73.7%, 60.5%, 13.2%; the conventional RT-PCR were 71.1%, 55.3%, 13.2%. There were no significant differences between the two methods.</p><p><b>CONCLUSION</b>The real-time fluorescent PCR detecting method of EV, EV71 and Cox A16 nucleic acid have been established successfully.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , DNA Primers , Chemistry , Genetics , Enterovirus A, Human , Genetics , Fluorescent Dyes , Chemistry , Hand, Foot and Mouth Disease , Diagnosis , Pathology , Virology , Polymerase Chain Reaction , MethodsABSTRACT
<p><b>OBJECTIVE</b>To demonstrate molecular characterization of a newly isolated enterovirus.</p><p><b>METHODS</b>Virus were isolated from patient with unknown-causing disease and tested by reverse transcription-polymerase chain reaction (RT-PCR) and 5'3'RACE (rapid amplification of cDNA ends, RACE), in an attempt to obtain the sequence of this newly isolated enterovirus.</p><p><b>RESULTS</b>Sequence analysis showed that this newly isolated enterovirus shared 83%-94% nucleotide identity and 91%-100% amino acid identity with enterovirus 89. Phylogenetic analysis indicated that it was probably a new subtype of enterovirus 89.</p><p><b>CONCLUSION</b>This newly isolated enterovirus in the stool specimen from patient has the same serotype with enterovirus 89, but it was probably a new subtype of enterovirus 89.</p>