No abstract available.
Antigens, Bacterial/*analysis ; Feces/*microbiology ; Helicobacter Infections/*diagnosis ; Helicobacter pylori/*isolation & purification ; Humans ; Immunoenzyme Techniques

Nine proteins encoded by Mycobacterium tuberculosis RD1 region are important protective antigens that become absent in long passaging of Mycobacterium tuberculosis. They only exist in pathogenic Mycobacteria and are absent in Bacille Calmette-Guerin and environmental Mycobacteria. With good immunogenicities, they may play an important role in the diagnosis and prevention of Mycobacterium tuberculosis. This article reviews recent studies on using RD1-encoded proteins as antigens in the diagnosis of active tuberculosis and tuberculous pleurisy.
Antigens, Bacterial ; isolation & purification ; metabolism ; Humans ; Mycobacterium tuberculosis ; physiology ; Tuberculosis ; diagnosis

Antibodies, Bacterial ; blood ; Antigens, Bacterial ; biosynthesis ; Bacterial Proteins ; biosynthesis ; Carrier Proteins ; biosynthesis ; Gastritis ; microbiology ; Helicobacter Infections ; immunology ; Helicobacter pylori ; immunology ; isolation & purification ; Humans ; Recombinant Proteins ; biosynthesis

OBJECTIVETo evaluate the accuracy of the Helicobacter pylori (H. pylori) stool antigen (HpSA) test and ImmunoCard STAT HpSA in the primary diagnosis of H. pylori infection.

METHODSWe searched Medline (1966-2007.4), EMbase (1985-2007.4), Chinese Journals Full-text Database (CJFD) (1994-2007) etc. to identify Clinical Trials of ImmunoCard STAT HpSA for the primary diagnosis of H. pylori infection. Meta-analysis was conducted using the method recommended by The Cochrane Collaboration Center.

RESULTSEleven trials were included with pooled sensitivity, pooled specificity as 0.93 (95% CI: 0.91-0.94), 0.93 (95% CI: 0.90- 0.95), respectively. Pooled positive likelihood ratio and pooled negative likelihood ratio were 12.01 (95% CI: 8.90-16.19), 0.08 (95% CI: 0.07-0.11), respectively with the pooled diagnostic odds ratio as 160.14(95% CI :100.43-255.34). The area under the summary receiver operating characteristic (SROC) was 0.974 +/- 0.005.

CONCLUSIONImmunoCard STAT HpSA appeared to be an accurate non-invasive method for the initial diagnosis of H. pylori infection.

Antigens, Bacterial ; immunology ; Feces ; microbiology ; Helicobacter Infections ; diagnosis ; immunology ; Helicobacter pylori ; immunology ; isolation & purification ; pathogenicity ; Reagent Kits, Diagnostic ; Sensitivity and Specificity

OBJECTIVETo analyze the genetic differences of Orientia tsutsugamushi (Ot) Sta56 gene between Shandong isolates and other strains deposited in GenBank.

METHODSPCR and restriction fragment length polymorphism (RFLP) were used to amplify the complete sequence of Ot-Sta56 gene. RFLP profiles of Ot were predicted by a computer program according to their complete sequences of Ot-Sta56 gene. PCR amplicon from XDM2 strain was sequenced and analyzed by Clustal X (1.8) and PHYLIP software.

RESULTSThe complete sequences (about 1.6 kbp) of Ot-Sta56 gene were amplified from B16 strain (isolated from patients), FXS2 strain (isolated from A. agrarius) and XDM2 strain. Four species of restriction endonucleases (Hha I, Hinf I, Hae III, Pst I) were used to digest the PCR amplicons from the 3 isolates. When comparing with the RFLP profiles of prototype Ot, the RFLP profiles of PCR amplicons from the 3 isolates were similar to those of Japan Kawasaki strain, but were quite different from the international reference strains Gilliam, Karp, Kato. Results from DNA sequence analysis showed that the complete sequence of Ot-Sta56 gene homology to Japan Kawasaki strain of XDM2 strain was 97%, and deduced amino acid sequence was 92%.

CONCLUSIONData from the complete sequence of Sta56 gene indicated that the genotypes of Ot isolates in Shandong province were similar, but with distinction from the Kawasaki strain.

Amplified Fragment Length Polymorphism Analysis ; Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; Bacterial Typing Techniques ; classification ; DNA, Bacterial ; genetics ; Genes, Bacterial ; Membrane Proteins ; genetics ; Orientia tsutsugamushi ; genetics ; isolation & purification ; Polymorphism, Restriction Fragment Length ; Sequence Analysis, DNA

Antigens, Bacterial ; immunology ; Antigens, Surface ; immunology ; China ; epidemiology ; Diarrhea ; microbiology ; Escherichia coli Infections ; epidemiology ; microbiology ; Escherichia coli O157 ; classification ; immunology ; isolation & purification ; Humans

OBJECTIVETo establish a recombinant plasmid of CFP32 of Mycobacterium tuberculosis in E. coli, and to analyze its antigenicity.

METHODSRv0577 gene was amplified by polymerase chain reaction from genome of Mycobacterium tuberculosis, and then cloned into vector pMD18-T followed by the subclone into the expression vector pET21a. Recombinant CFP32 was expressed and purified. The antigenicity of the recombinant protein was analyzed by using Western-blot. The purified recombinant CFP32 protein was used as an antigen to screen the sera of 7 pulmonary TB patients (n = 97), as well as the other pulmonary disease patients (n = 25), and the clinically healthy controls (n = 38) by ELISA.

RESULTSRecombinant plasmid of CFP32 was established, and be expressed efficiently in E. coli BL21 (DE3). The relative molecular mass of the protein was about 300,000 by SDS-PAGE analysis. The protein purified by Ni-NTA was in a purity over 90%, which was confirmed by Western-blot analysis. ELISA analysis showed its sensitivity and specificity were 63.9% (62/97) and 96.8% (2/63) respectively.

CONCLUSIONThe recombinant expression plasmid pET21a CFP32 has been constructed and CFP32 proteins has been successfully expressed and be purified in E. coli and, ELISA analysis has identified the recombinant CFP32 as a candidate antigen for TB serodiagnosis.

Antigens, Bacterial ; blood ; Bacterial Proteins ; genetics ; immunology ; Cloning, Molecular ; Escherichia coli ; Gene Expression ; Humans ; Mycobacterium tuberculosis ; genetics ; isolation & purification ; Plasmids ; Recombinant Proteins ; Serologic Tests ; Tuberculosis, Pulmonary ; diagnosis ; microbiology

To set up a method of amplification for the whole CagA gene of Helicobacter pylori and its fingerprinting by restriction fragment length polymorphism (RFLP), nested PCR was employed in combination with TD-PCR to amplify the gene and EcoRI and Hind III were used to generate the RFLP fingerprinting. Target DNA fragments from 13 of 20 samples were successfully amplified and the relevant RFLP fingerprintings were obtained. It is concluded that the method can be used to amplify the whole CagA gene and CagA gene has apparent diversity of RFLP profile.
Antigens, Bacterial ; genetics ; Bacterial Proteins ; genetics ; DNA Fingerprinting ; methods ; Gene Amplification ; genetics ; Helicobacter pylori ; genetics ; isolation & purification ; Humans ; Polymorphism, Restriction Fragment Length

We utilized polyethylenimine (PEI) and Glu solution to immobilize the antibody of Treponema Pallidum on the silver electrode of piezoelectric quartz crystal and detect the standard antigen solution at different concentration. The antibody keeps the intrinsic Y type molecular structure revealed by AFM. The resonant range of the sensor is between 5 x 10(-5) - 1.25 x 10(-4) g/mL and the correlation coefficient is 0.9976, the optimal resonance pH is 7.5. We found the sensor having good selectivity in comparison with the method using BSA. At last, a discussion on the reproducibility of the sensor is presented.
Antibodies, Bacterial ; immunology ; Antigens, Bacterial ; analysis ; Biosensing Techniques ; instrumentation ; Equipment Design ; Immunoassay ; instrumentation ; methods ; Microelectrodes ; Polyethyleneimine ; Quartz ; Treponema pallidum ; immunology ; isolation & purification

OBJECTIVETo understand the distribution of emm gene types related to group A streptococcus-caused scarlet fever among children in Beijing and to analyze the relationship between the mutation of the emm types and scarlet fever.

METHODSNasopharyngeal swab samples were collected from the scarlet fever cases diagnosed in 36 hospitals in Beijing to isolate the GAS strains from May to July, betgween 2011 and 2014. Genotyping of emm gene was performed with PCR and N-terminal gene fragments of M protein were sequenced. Data of all the scarlet fever cases in Beijing that reported through the National Notifiable Infectious Disease Surveillance System (NNIDSS) , were gathered and analyzed.

RESULTSAmong the collected 2 161 nasopharyngeal swabs, 762 GAS strains were identified (35.3%). In addition, 7 emm types were detected, in which emm12 accounted for 69.4% (529/762) , emm1 accounted for 29.8% (227/762) , and other five types (emm 11, 22, 75, 89, and 128) accounted for 0.8% (6/762) , respctively. Compared with the emm types detected between 2011 and 2014, emm12, emm1 and other types accounted for 82.2% (295/359) , 16.7% (60/359) and 1.1% (4/359, including emm11, 22 and 89) in 2011 respectively.emm12, emm1 and emm75 accounted for 77.3% (123/163) , 23.9% (39/163) and 0.6% (1/163) respectively in 2012. emm12 and emm1 accounted for 50.7% (38/75) and 49.3% (37/75) in 2013 while emm12, emm1 and emm128 accounted for 44.2% (73/165) , 55.2% (91/165) and 0.6% (1/165) respectively in 2014. The differences of the constitution of emm types from 2011 to 2014 appeared statistically significant (P<0.001). In 2011 and 2012, major type appeared as emm12, but in 2014, emm1 became predominant. A total of 6 152 cases were reported in 2011, while 2 908, 2 048 and 3 918 cases were reported in 2012, 2013 and 2014 respectively. Age specific differences were noticed in the distribution of emm types GAS strains in 2011, with the number of emm12 strains detected higher in 1-5 year olds than in age group > 5 years (P<0.05). There were area specific differences in distribution of emm types of GAS strains seen in 2011 and 2013. In 2011, the number of emm1 strains detected in urban area was higher than in suburb area (P<0.05). However, in 2013, the number of emm1 strains detected in suburb area was seen higher than in urban area (P< 0.05).

CONCLUSIONGAS with emm12 and GAS emm1 appeared interchangeably predominant in Beijing from 2011 to 2014. Changes in predominant emm types seemed also related to the trends of incidence rates on scarlet fever.

Antigens, Bacterial ; Bacterial Outer Membrane Proteins ; Beijing ; epidemiology ; Carrier Proteins ; Child ; Genotype ; Humans ; Mutation ; Scarlet Fever ; epidemiology ; genetics ; Streptococcus pyogenes ; classification ; genetics ; isolation & purification