[Show abstract][Hide abstract] ABSTRACT: Coxsackievirus A8 (CV-A8), a member of the genus Enterovirus of the family Picornaviridae, can cause a variety of infectious diseases, such as hand, foot and mouth disease (HFMD), herpangina (HA), encephalitis, paralysis, myelitis, and meningitis. This is a first report of complete genome sequences of CV-A8 strains associated with HFMD/HA since the prototype strain Donovan was identified in 1949. The complete genome sequences of eight new CV-A8 strains showed 19.2 %-20.6 % nucleotide differences when compared to the prototype strain Donovan, and 81.5 %-99.9 % similarity to each other. The topology of a polyphyletic tree based on complete capsid protein gene sequences indicated that the new CV-A8 strains and Donovan are monophyletic. However, seven CV-A8 strains clustered with CV-A10 and CV-A2 in the 5'UTR and P2 region, respectively. In the P3 region, three and four CV-A8 strains grouped with CV-A6 and CV-A2, respectively. Seven CV-A8 strains segregated from Donovan and grouped in a separate lineage in the 3'UTR. The strain CVA8/SZ266/CHN/2014 was most similar to EV71 in the nonstructural proteins regions. Phylogenetic analysis classified worldwide CV-A8 isolates into four distinct clusters, and almost all Chinese and Thai CV-A8 strains evolved independently in their respective lineages, which indicated geographical evolution of CV-A8.
Archives of Virology 10/2015; DOI:10.1007/s00705-015-2646-1 · 2.39 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: This is a report of the complete genomic sequences of two rare group C rotavirus strains RVC/SZ94/CHN/2011 and RVC/SZ272/CHN/2011,
isolated from two cases of acute gastroenteritis in Shenzhen, southern China, in 2011. These two strains display a close genetic
relationship to 2007 Chinese strain YNR001 and 2008 Japanese strain BK0830.
[Show abstract][Hide abstract] ABSTRACT: We determined the complete genome sequence of a coxsackievirus A16 strain (CVA16/SZ29/CHN/2014) from a fatal case in Shenzhen,
southern China, in 2014. The strain was assigned to subgenotype B1b based on phylogenetic analysis of the VP1 gene.
[Show abstract][Hide abstract] ABSTRACT: An outbreak of norovirus GII.4/Sydney_2012 affected a China elder care facility in December 2012. A total of 39 elderly people and staff met the outbreak case definition. The attack rates in the elderly and the staff were 15.9% (31/195) and 23.2% (19/82), respectively, including 13 asymptomatic cases in the staff. The result of gene sequencing revealed that the outbreak was caused by norovirus GII.4 Sydney. The mode of transmission of this outbreak was proven to be person-to-person. The first case (a self-cared elder) was affected outside the elder care facility and was not isolated after returning. Norovirus was transmitted via close contact among the self-cared elderly. Then, through service-related close contact, the attendants promoted the cross-transmission between the self-cared elderly and the nursed elderly. The virus was also spread among the staff via daily contact. In the elder care facility, the asymptomatic cases in the attendants played an important role in the transmission of norovirus, which deserves high attention.
Japanese journal of infectious diseases 02/2015; 68(1):70-4. DOI:10.7883/yoken.JJID.2014.081 · 1.16 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Viral gastroenteritis is one of the most common diseases in humans, and it is primarily caused by rotaviruses (RVs), astroviruses (AstVs), adenoviruses (AdVs), noroviruses (NoVs), and sapoviruses (SaVs). In this study, we determined the distribution of viral gastroenteritis and human calicivirus (HuCVs) in acute gastroenteritis patients in Shenzhen, China, during 2011. Real-time RT-PCR was used to detect norovirus (NoV), group A rotavirus (RV), adenovirus (AdV), and astrovirus (AstV). From a total of 983 fecal samples, NoV was detected in 210 (21.4 %); RoV in 173 (17.6 %); AstV in 10 (1.0 %); and AdV in 15 (1.5 %). Mixed infections involving two NoVs were found in 21 of the 387 pathogen-positive stool specimens. NoV and SaV genotypes were further tested using RT-PCRs and molecular typing and phylogenetic analysis were then performed based on the ORF1-ORF2 region for NoV and a conserved nucleotide sequence in the capsid gene for SaV. Of the 68 typed strains that were sequenced and genotyped, five were NoV G1 (7.5 %) and 63 were NoV GII (96.6 %). GII strains were clustered into five genotypes, including GII.4 (65.1 %; 36 GII.4 2006b and five GII.4 New Orleans), GII.3 (28.6 %), GII.2 (3.2 %), GII.6 (1.6 %), and GII.1 (1.6 %). While all fecal specimens were tested for SaVs, 15 (1.5 %) were positive, and of these, 12 isolates belonged to G1.2, and the remaining three SaV strains belonged to the SaV GII genogroup. Although various HuCVs were detected in acute gastroenteritis patients, NoV GII.4 2006b was more prevalent than the other HuCVs.
Archives of Virology 03/2014; 159(8). DOI:10.1007/s00705-014-1986-6 · 2.39 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Sporadic hand, foot and mouth (HFMD) outbreaks and other infectious diseases have frequently been associated with certain human enterovirus (HEV) serotypes in recent years. This study explored the prevalence and genetic characteristics of non-HEV71 and non-CV-A16 human enteroviruses associated HFMD infections in Shenzhen, China. A total of 2411 clinical stool specimens were collected from a hospital-based surveillance for HFMD from 2008 to 2012. The detection of HEV was performed by real-time RT-PCR and RT-seminested PCR, and spatiotemporal phylogenetic analysis was performed based on the VP1 genes. A total of 1803(74.8%) strains, 28 different serotypes were detected. In the past 5 years, the predominant serotype was HEV71 (60.0%), followed by CV-A16 (21.2%) and two uncommon serotypes CV-A6 (13.0%) and CV-A10 (3.3%). However, CV-A6 replaced CV-A16 as the second most common serotype between 2010 and 2012. Being an emerging pathogen, CV-A6 turned into an equally common causative agent of HFMD as HEV71 in Shenzhen, 2012. Phylogenetic analysis revealed that little variation occurred in Chinese HEV71 and CV-A16 strains. The genetic characteristics of Chinese CV-A6 and CV-A10 strains displayed geographic differences. CV-A6 and CV-A10 circulating in Shenzhen likely originated from European countries. Human enteroviruses revealed a rapid mutation rate due to evolutionary pressure and frequent recombination (3.2×10(-3)-6.4 ×10(-3) substitutions per site per year for HEV71, CV-A6, CV-A16 and CV-A10). Since certain serotypes are potential threatens to the public health, this study provides further insights to the significance of epidemiological surveillance of HFMD.
[Show abstract][Hide abstract] ABSTRACT: To determine the prevalence and distribution of human enteroviruses (HEVs) among healthy children in Shenzhen, China.
Clinical specimens were obtained from 320 healthy children under 5 years old in Shenzhen, China from 2010 to 2011. The specimens were evaluated using real-time PCR and cell cultures. The positive specimens were further tested using reverse transcription-seminested PCR (RT-snPCR). Molecular typing and phylogenetic analysis were based on the sequence determined.
Among the 320 samples, 34 were tested positive for HEVs (10.6%) and 22 different serotypes were identified using RT-snPCR. PV1 and PV2 were also detected. The predominant serotype observed was EV71 (17.6%), followed by CV-B4 (14.7%). HEV-B was detected most frequently, with an overall prevalence of 47.1%. HEV-A and HEV-C were found in 32.3% and 20.6% of the samples, respectively. No HEV-D was identified. Molecular phylogeny indicated that all EV71 strains were of C4 genotype.
Although a variety of HEVs was detected in healthy children, HEV-B was relatively more prevalent than other HEV species. Considering HEV-A is more prevalent than HEV-B among patients with hand-foot-mouth disease, additional long-term surveillance of HEV is warranted in both asymptomatic and symptomatic populations.
PLoS ONE 06/2013; 8(6):e64889. DOI:10.1371/journal.pone.0064889 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To obtain information on viral molecular structural and evolutionary characteristics, we conducted the SZ2010422 full-length genomic analysis.
Primers were designed by New Orleans full sequence, SZ2010422 full genome was amplified by RT-PCR, the whole genome sequence and the capsid domain amino acid sites was analysised after cloned and sequenced.
The genome of G II-4 Norovirus SZ2010422 strain was consist of 7559 bp, it revealed three ORFs composites of the whole genome, ORF1 (5100 bp), ORF2 (1623 bp), ORF3 (807 bp) respectively, ORF1 and ORF2 had 19 nucleotide overlap. By evolutionary comparative analysis found SZ2010422 genomic nucleotide sequences with reference strains of G II-4 New Orleans1805 strains the highest homology with a total length of homology was 99.3%, of ORF1 (99.5%), ORF2 (99.2%), ORF3 (98.6%). Phylogenetic analyses showed SZ2010422 belonging to G II-4 New Orleans variant. Date of 541 amino acid analyses showed: New Orleans variant strains of popular sites: aa310N or K, --> S aa341D --> of N, aa359T--> S, aa396H --> P, aa460H --> Y.
Norovirus SZ2010422 belonged to the G II-4 New Orleans variant. In This study, SZ2010422 full sequence can be used not only as a full-length NoV variant sequence standard for future comparison studies, but also as useful material for the public health field by enabling the diagnosis, vaccine development, and prediction of new emerging variants. Noroviruses; Genes; Sequence analysis
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 06/2013; 27(3):181-3.
[Show abstract][Hide abstract] ABSTRACT: To analyze the genetic evolution for the common causative agent of hand, foot and mouth disease(HFMD) that VP4 of human enterovirus 71 in Shenzhen district.
491 sttol specimen were collected from, children with hand, foot and mouth diease in Shenzhen Children's Hospital 2009. After cell culture, VP4 gene of eight EV71 strains were amplified by reverse-transcriptase PCR( RT-PCR), phylogenetic analysis of the VP4 gene was constructed by using MEGA 4. 0.
The VP4 gene of eight EV71 strains encoding 69 amino acids with full length 207 bp. The nucleotide homology of VP4 gene among eight EV71 strains was 94. 2% -98. 1%, compared with VP4 gene of EV71 strains retrieved from Shenzhen 2001 to 2004 and GenBank was 89. 1%-98. 1% and 79.2%-100% respectively. Asian epidemic strain Fuyang had the highest nucleotide homology, representative strain C4 and Shenzhen strain (AY895144) with 94. 2% -98. 1% secondly. Except for the 54th amino acid of VP4 gene of India reported strain and one of the eight EV71 strains, the homology of the rest amino acids between the eight EV71 strains and those in GenBank was 100%. Compared with representative strain C4,there were seventeen differences in nucleotide sequences of VP4 of the eight EV71 strains. All of the different nucleotides were located at the degenerate password sites except one. There was no significant difference in VP4 gene between the severe and the mild cases of strainS. The eight Shenzhen EV71 strains were classified as sub-genotype C4 in the phylogenetic tree.
The epidemic of EV71 in Shenzhen 2009 was sub-genotype C4. VP4 gene of EV71 was very conservative which dose not belong to the variation section. The variation of most of nucleotide was invalid variation. The amino acids encoded by VP4 gene which variation was almost zero.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 02/2013; 27(1):11-3.
[Show abstract][Hide abstract] ABSTRACT: To describe epidemiologic characteristics of norovirus infection and its genotype in Shenzhen area of 2010. Stool specimens were collected from four monitoring point and detected by reverse transcription polymerase chain reaction (PT-PCR). Positive PCR products were purified and sequenced, and the sequences were performed by Clustal W and MEGA 4.0 programs, then genotype was identified and phylogenetic tree was constructed. Nucleotide sequence analysis revealed that 79 strains of NV belonged to norovirus genogroup II and 6 belonged to genogroup 1 of all 85 positive products. While 55 strains belonged to G II/4(2006b), 16 strains belonged to G II/4(2008 variant), 2 strains belonged to G II /1, 4 strains belonged to G II/5, 2 strains belonged to G II/11, 1 strain belonged to GII/4, 2 strains belonged to GII/5, 3 strains belonged to GI/6. The main genogroups of norovirus in Shenzhen ware GI and G II. G II /4 was one of the most major genotype of norovirus , while G II /4(2006b) variant was identified as the predominant strain in Shenzhen area.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 05/2012; 28(3):219-23.
[Show abstract][Hide abstract] ABSTRACT: To conduct an epidemiological and genotype analysis of sapovirus (SaV) associated with sporadic diarrhea in Shenzhen in the year 2009.
A total of 852 fecal samples were collected from sporadic cases of diarrhea in Shenzhen in 2009 and detected by reverse-transcription polymerase chain reaction (RT-PCR) using the primers of SLV5317/5749. The PCR products were analyzed with 1.5% agarose gel electrophoresis and sequenced to construct the phylogenetic tree.
Sixteen samples were found positive for SaV, with a positivity rate of 1.88%. Sequence analysis identified 8 isolates as SaV GI genotype (including 3 SaV GI.1 and 5 SaV GI.2), 7 as SaV GIV genotype, and 1 as GII genotype.
SaV infection is present in Shenzhen with GI as the predominant genotype. This is the first report of SaV GIV strains in China, which differs from the strains of Anhui-A141 and Beijing-CHN99/BJ360, suggesting the genotypic variety of SaV infection in China.
Nan fang yi ke da xue xue bao = Journal of Southern Medical University 06/2011; 31(6):1017-9.
[Show abstract][Hide abstract] ABSTRACT: Genetic evolution of VP1 of enterovirus type 71 in Shenzhen were analyzed.
All samples were tested by RT-PCR using EV71 specific primer. The VP1 of EV71 were amplified and sequenced. A phylogenetic tree was constructed by comparison of the sequences with subgenotype A, B and C using DNAStar, BioEdit and Mega 3.1 software.
Among 35 strains, the homogeneity of the VP1 nucleotide sequence was between 92.1%-100%. The homogeneity of the VP1 nucleotide sequence with subgenotype A and B was between 81.4% -91.1%. The VP1 nucleotide sequence of 35 strains of Shenzhen shared between 93% -97.4% identity with cluster C4. The prevalence strains of EV71 were cluster C4b from 1998 to 2004, and gradually moved to C4a since 2003. All of EV71 were C4b from 2006 to 2008. Also, the homogeneity of the VP1 nucleotide sequence with Anhui FY23 EV71 strain were 94.5% -94.7%, 95.7% -95.8%, 96.2%, 95.4% -97.5%, 96.3% -99.2% from 2003 to 2008. It shows that the homogeneity was increased year by year. There was a mutation (A --> C) at No. 66 nucleotide of VP1 of EV71 that two strains were isolated in 2003 and 8 strains in 2008, that caused amino acid mutation (Q --> H) at No. 22 of VP1.
EV71 C4b was gradually moved to C4a from 1998 to 2008. There was a missense mutation at No. 66 nucleotide of VP1.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 06/2011; 25(3):173-5.
[Show abstract][Hide abstract] ABSTRACT: Noroviruses (NoVs) cause gastroenteritis in all age groups. To investigate an acute gastroenteritis outbreak in Shenzhen,
South China, we analyzed the presence of NoVs in fecal specimens and drinking water samples. Viral RNAs were extracted from
fecal suspensions and water concentrates made by membrane adsorption-elution, and NoV was detected by reverse transcription-polymerase
chain reaction (RT-PCR). Among the 43 diseased workers at the two factories and one construction site, NoVs were detected
in 19 out of 28 stool specimens (68%) and 3 out of 16 water samples (19%). Importantly, the six NoV strains identified from
the water and the stool samples share over 99% sequence identity and all belong to GII 4 viruses. Epidemiological survey also
identified a water pipe breakage possibly causing the water contamination, and confirmed that the contaminated drinking water
largely contributed to the NoV gastroenteritis outbreak. To our knowledge, this is the first time that a viral concentration
and detection method for waterborne NoV is applied to a gastroenteritis outbreak in China.
KeywordsNorovirus-Acute gastroenteritis-Norovirus outbreak-Water-Genotype-PCR
[Show abstract][Hide abstract] ABSTRACT: To understand the immunological status of Japanese encephalitis (JE) antibodies amongst migrant workers and to provide epidemiological basis for public health strategies on JE prevention and control in Shenzhen.
A multi-stage random sampling method was used, and 1003 migrant workers aged 18 to 60 from 44 factories were investigated and their serum specimens were collected. The enzyme-linked immunosorbent assay (ELISA) was used to detect JE antibodies qualitatively.
The gross IgG seroprevalence rate for JE was 20.2% (203/1003). Sex-specified seroprevalence was 21.2% (103/485) for male and 19.3% (100/518) for female, respectively (χ(2) = 579, P > 0.05). Age-specific seropositive rates were 22.6% (12/53) for those below 20 years old, 18.7% (120/642) for those between 20-years old, 26.0% (58/223) for those between 30-years old and 15.3% (13/85) for those on or above 40 years old (χ(2) = 7.96, P > 0.05). Proportions for self-reported positive immunization, non-immunization and unclear immunization history were 22.1% (30/136), 22.1% (51/231) and 19.2% (122/636), respectively (χ(2) = 501, P > 0.05). Seroprevalence by region of origins showed that workers from Guangdong province was the highest (30.5%, 50/164), followed by workers from Guangxi (29.7%, 22/74) whilst workers from Shan(3)xi (5.4%, 2/37) had the lowest rate. Seroprevalence rate for managers (29.0%, 31/107) was higher than that of technicians (7.1%, 1/14) (χ(2) = 21.78, P < 0.05). Serological positive rate of workers with university or above educational background was the highest (32.7%, 16/49), followed by that for individuals with college degree (10.3%, 10/97) (χ(2) = 13.02, P < 0.05).
No associations are detected between JE seroprevalence and age, or sex, or self-reported immunization histories amongst migrant labor workers in Shenzhen. However, correlations between JE serological positive rate and region of origins, occupation and educational attainment are found to be significant. The gross seroprevalence of JE antibodies suggests that the level of JE antibodies amongst Shenzhen migrant workers is low and the population immunity barrier has yet to be established. It is necessary to strengthen prevention and control strategies of JE among labor workers of Shenzhen.
Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] 09/2010; 44(9):806-9.
[Show abstract][Hide abstract] ABSTRACT: To study the binding profile of NV strain SZ9711 (GII-4) with human histo-blood group antigens (HBGAs).
The P domain-encoding fragment was amplified by RT-PCR from the stain SZ9711 and cloned into the pGEX-4T-1 vector. The recombinant fusion protein was expressed in E. coli and purified using the column Sepharose 4B. The P protein was released by thrombin cleavage. The binding of P particles of SZ9711 and VA387 with the HBGAs were measured by saliva-based EIA method.
The expression of the recombinant fusion protein was shown by the SDS-PAGE, in which a 38 x 10(3)-P protein was obtained. Saliva-based EIA revealed that the P particle of SZ9711 bound to HBGAs in saliva similar to that of the strain VA387 reported previously. It bound strongly to saliva of type A, B and O(secretor) but did not interact with saliva of type O(non-secretor). Noteworthy, binding ability of SZ9711 P particle to type A saliva was lower than that of the VA387 P particle.
This is the first time that a P particle was prepared from a norovirus strain isolated in China and the binding ability of the P particle with HBGAs was analyzed. The result indicated the binding profile of the SZ9711 P particle was similar to that of VA387 reported previously. These data may be valuable in studying the relationship between noroviruses and their bindings to HGBA receptors.
Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology 02/2010; 24(1):5-7.
[Show abstract][Hide abstract] ABSTRACT: Genetic characteristics of enterovirus type 71 in Shenzhen from 2005 to 2008 were analyzed. All samples were detected by RT-PCR using EV71-specific primers. The VP1s of EV71 strains were amplified and sequenced. A phylogenetic tree was constructed by comparison of the sequences with those of subgenotype A, B and C using DNASTAR, BioEdit and Mega 3.1 softwares. The VP1 nucleotide sequences of 17 strains of Shenzhen shared 95.3%-99.4% identities with cluster C4a, and one strain shared 93.0%-95.6% identities with cluster C4b. The homogeneity of the VP1 nucleotide sequence with subgenotype A and B was 81.8%-86.0%. Among 18 strains, the homogeneity of the VP1 nucleotide sequence was between 92.5%-100%. All EV71 strains circulating in Shenzhen were of subgenotype C4. The predominant strain of EV71 belonged to cluster C4a, also there was cluster C4b.
Bing du xue bao = Chinese journal of virology / [bian ji, Bing du xue bao bian ji wei yuan hui] 11/2009; 25(6):415-9.
[Show abstract][Hide abstract] ABSTRACT: To isolate and identify the pathogen of Dengue fever from Shenzhen city in 2005 - 2006, and to analyze the molecular characteristics of the isolated Dengue virus strain as well as to explore its possible origin.
IgM and IgG of serum samples taken from 60 suspected Dengue fever patients were detected by ELISA and immunochromatography, and 9 specimens were positive. Nine samples from patients with early stage Dengue fever were used to isolate virus with C6/36 cell line and the positive cell cultures were identified by MGB fluorescent PCR. The type of isolated virus strain was determined by RT-semi-nested-PCR and fluorescent PCR. E gene of isolated virus strain was amplified by RT-PCR and sequenced. Homology and phylogenetic tree of E gene of Shenzhen Dengue virus with the strains isolated from other areas were constructed.
Of nine antibody-positive serum samples, one strain of Dengue virus was successfully isolated. The isolated virus strain was confirmed as Dengue virus type 2 and designated as DEN2-SZ0521. The homology of nucleotide sequence and the deduced amino acid sequence of E gene of SZ0521 with standard type 2 Dengue virus NGC strain was 94.2% and 98.2%, but the homology with standard Dengue virus 1, 3, 4 in the same fragment were 59.1%, 57.2%, 58.5% and 68.1%, 66.7%, 63.2%, respectively. The phylogenetic tree indicated that SZ0521 had the greatest similarity with the Malay0412a/Tw strain and they lied in the same branch of the phylogenetic tree. The corresponding homology of nucleotide sequence and amino acid sequence was 99.8% and 100%, respectively. The isolated Dengue virus type 2 belonged to genotype IV with Indonesia-76, Somalia-84 and Sri Lanka-90.
Dengue virus was isolated from Shenzhen for the first time, and it was classified as type 2. It was confirmed that the type 2 Dengue virus may come from the epidemic area in Malaysia.
Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine] 09/2009; 43(9):798-802.
[Show abstract][Hide abstract] ABSTRACT: To carry out genotype analysis of enterovirus type 71, detected from hand-foot-mouth disease patients in Shenzhen in 2004.
All samples were tested by reverse transcription-polymerase chain reaction (RT-PCR) using EV71 specific primer. The VP1 and VP4 of EV71 were cloned and sequenced. A phylogenetic tree was constructed by comparing the sequences with genogroups A, B and C genotypes using TreeView and PHYLIP software (3.6b).
The VP1 nucleotide sequence of 4 strains isolated from Shenzhen shared 87.8% - 92.0% identity with genogroup C, while the homogeneity of the VP4 nucleotide sequence was between 85.9% - 87.4%. The homogeneity of the VP1 nucleotide sequence with genotypes A and B was between 81.9% - 84.2% and was 80.6% - 85.0% with VP4. Among the 4 strains, the homogeneity of the VP1 nucleotide sequence was between 94.1% - 99.8% and was 100% with VP4 which formed a small group and could denominate EV71 genetype C4.
Similar results were obtained from phylogenetic analysis of EV71 based on VP1 and VP4 nucleotide sequence. The four EV71 strains causing epidemic in Shenzhen could be named as C4 subgroups.
Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi 09/2008; 29(8):790-3.
[Show abstract][Hide abstract] ABSTRACT: To develop the Taqman MGB real-time fluorescent PCR assay for rapid detection of Dengue viruses.
Using Taqman MGB technique, a pair of universal primers and Taqman MGB probe were designed according to a highly reserved sequence of the 3'-noncoding region of dengue viruses type 1-4. Dengue virus strains were used as standard and Japanese encephalitis virus strains were used as control, the real-time PCR assay for specific and sensitive detection of the dengue viruses was established. While 8 serum specimens of ELISA positive were detected by the RT-PCR and fluorescent PCR.
The sensitivity of real-time PCR was 0.17pg/microl (cDNA)or 10(-5)TCID50. There was no cross-reaction with Japanese encephalitis virus. Of 8 specimens, 2 were positive by RT-PCR and 5 were positive by real-time PCR. The test could be completed in 4 hours.
The Taqman MGB real-time PCR assay was fast, sensitive and specific. It could be applied to the quick early diagnosis of dengue viruses.
Wei sheng yan jiu = Journal of hygiene research 12/2006; 35(6):736-8.