Objective: To develop a sensitive, specific and simple method for detection of extremely low numbers of T. pallidum in clinical specimens, as a significant addition to the serologic tests for syphilis diagnosis. Metho...Objective: To develop a sensitive, specific and simple method for detection of extremely low numbers of T. pallidum in clinical specimens, as a significant addition to the serologic tests for syphilis diagnosis. Methods: Double-tube nested PCR(DN-PCR) and single-tube nested PCR(SN-PCR) assays were performed to amplify specific fragments of the DNA poly-merase I gene(polA) of T. pallidum. Sensitivity and specificity of the two PCR assays were tested. Eighty-six whole blood specimens from persons with suspected syphilis were detected by the two nested PCR methods. The TPPA test was used as a comparison for detecting syphilis in sera from corresponding patients. Results: Only specific amplicons could be obtained during amplification of the T. pallidum polA gene and the detection limit was approximately 1 organism when analyzed on gel by the two PCR methods. Of 86 clinical specimens, 62 were positive by TPPA. Of these, 54 and 51 were positive by the DN-PCR and SN-PCR, respectively, which does not represent a statistically significant difference between the two PCR tests. Of 24 TPPA-negative specimens, 5 were positive by both DN-PCR assay and SN-PCR assay. Conclusion: The SN- polA PCR method is extremely sensitive, specific and easy to perform for detecting low numbers of T. pallidum in clinical blood specimens as a complementary to serology for syphilis diagnosis.展开更多
Background： Direct ZN （Ziehl-Neelsen） sputum smear microscopy for diagnosis of TB （tuberculosis） has low sensitivity, especially in TB/HIV co-infected patients. Sputum concentration by bleach （NaOCI） with sedim...Background： Direct ZN （Ziehl-Neelsen） sputum smear microscopy for diagnosis of TB （tuberculosis） has low sensitivity, especially in TB/HIV co-infected patients. Sputum concentration by bleach （NaOCI） with sedimentation has been used to increase the sensitivity of sputum smear microscopy in many settings but with varying results. Objective： To determine whether bleach plus centrifugation significantly improves the detection of AFB （acid-fast bacilli） in ZN smear-negative sputum specimens. Methods： Three hundred and seventy sputum specimens were collected from new TB suspects attending a Nairobi referral district hospital and processed for direct microscopy using ZN technique and culture on Lowenstein Jensen Media. All smear-negative specimens were treated with 3.5% bleach and left to stand for 30 min before centrifugation. The bleach treated smears were processed and examined using ZN technique. Results： Of the 370 specimens, 200 （54%） were positive culture. The number of sputum samples that were smear-positive by direct ZN was 138 （37.2%）, with a sensitivity of 66%. After treatment of direct ZN smear-negative specimens with 3.5% bleach and centrifugation, the total number of AFB smear-positive samples increased to 171 with an increase in sensitivity of 66% to 81.1% （15.1%）. Conclusion： In this study, bleach with centrifugation significantly increased the yield of sputum smear microscopy. Further evaluation of these techniques in routine programmes is required especially in settings where the burden of TB/HIV is high.展开更多
CDC48 is a highly conserved protein in eukaryotes and belongs to the AAA （ATPase associated with a variety of cellular activities） superfamily. It can interact with many different cofactors and form protein complexe...CDC48 is a highly conserved protein in eukaryotes and belongs to the AAA （ATPase associated with a variety of cellular activities） superfamily. It can interact with many different cofactors and form protein complexes that play important roles in various cellular processes. According to the Physcomitrella patens database, one member of the ATPases, the cell cycle gene PpCDC4811, was cloned. PpCDC48II contains two typical ATPase modules and is highly homologous to AtCDC48A. PpCDC4811 was up-regulated in mRNA levels after incubation at 0~C for 36 and 72 h. To further elucidate protein function, we disrupted the PpCDC4811 gene by transforming P. patens with the corresponding linear genomic sequences. When treated to the same freezing stress, it was found that PpCDC4811 knockout plants were less resistant to freezing treatment than wild type after acclimation. This suggested that PpCDC481I was an essential gene for low-temperature-induced freezing tolerance in P. patens cells.展开更多
In this work,we parallelly detected the specific binding between microarray targets including 12 different kinds of proteins and the probe solution containing five corresponding antibodies and quantitatively analyzed ...In this work,we parallelly detected the specific binding between microarray targets including 12 different kinds of proteins and the probe solution containing five corresponding antibodies and quantitatively analyzed the interactions between CDH13 and solution phase anti-CDH13 at six different probe concentrations by oblique-incidence reflectivity difference(OIRD)method in label-free format.The detection sensitivity reached 10 ng/mL.The experimental results indicate that the OIRD method is a promising and competing technique not only in research work but also in clinic.展开更多
文摘Objective: To develop a sensitive, specific and simple method for detection of extremely low numbers of T. pallidum in clinical specimens, as a significant addition to the serologic tests for syphilis diagnosis. Methods: Double-tube nested PCR(DN-PCR) and single-tube nested PCR(SN-PCR) assays were performed to amplify specific fragments of the DNA poly-merase I gene(polA) of T. pallidum. Sensitivity and specificity of the two PCR assays were tested. Eighty-six whole blood specimens from persons with suspected syphilis were detected by the two nested PCR methods. The TPPA test was used as a comparison for detecting syphilis in sera from corresponding patients. Results: Only specific amplicons could be obtained during amplification of the T. pallidum polA gene and the detection limit was approximately 1 organism when analyzed on gel by the two PCR methods. Of 86 clinical specimens, 62 were positive by TPPA. Of these, 54 and 51 were positive by the DN-PCR and SN-PCR, respectively, which does not represent a statistically significant difference between the two PCR tests. Of 24 TPPA-negative specimens, 5 were positive by both DN-PCR assay and SN-PCR assay. Conclusion: The SN- polA PCR method is extremely sensitive, specific and easy to perform for detecting low numbers of T. pallidum in clinical blood specimens as a complementary to serology for syphilis diagnosis.
文摘Background： Direct ZN （Ziehl-Neelsen） sputum smear microscopy for diagnosis of TB （tuberculosis） has low sensitivity, especially in TB/HIV co-infected patients. Sputum concentration by bleach （NaOCI） with sedimentation has been used to increase the sensitivity of sputum smear microscopy in many settings but with varying results. Objective： To determine whether bleach plus centrifugation significantly improves the detection of AFB （acid-fast bacilli） in ZN smear-negative sputum specimens. Methods： Three hundred and seventy sputum specimens were collected from new TB suspects attending a Nairobi referral district hospital and processed for direct microscopy using ZN technique and culture on Lowenstein Jensen Media. All smear-negative specimens were treated with 3.5% bleach and left to stand for 30 min before centrifugation. The bleach treated smears were processed and examined using ZN technique. Results： Of the 370 specimens, 200 （54%） were positive culture. The number of sputum samples that were smear-positive by direct ZN was 138 （37.2%）, with a sensitivity of 66%. After treatment of direct ZN smear-negative specimens with 3.5% bleach and centrifugation, the total number of AFB smear-positive samples increased to 171 with an increase in sensitivity of 66% to 81.1% （15.1%）. Conclusion： In this study, bleach with centrifugation significantly increased the yield of sputum smear microscopy. Further evaluation of these techniques in routine programmes is required especially in settings where the burden of TB/HIV is high.
基金supported by the National Natural Science Foundation of China (Grant No. 30700404)
文摘CDC48 is a highly conserved protein in eukaryotes and belongs to the AAA （ATPase associated with a variety of cellular activities） superfamily. It can interact with many different cofactors and form protein complexes that play important roles in various cellular processes. According to the Physcomitrella patens database, one member of the ATPases, the cell cycle gene PpCDC4811, was cloned. PpCDC48II contains two typical ATPase modules and is highly homologous to AtCDC48A. PpCDC4811 was up-regulated in mRNA levels after incubation at 0~C for 36 and 72 h. To further elucidate protein function, we disrupted the PpCDC4811 gene by transforming P. patens with the corresponding linear genomic sequences. When treated to the same freezing stress, it was found that PpCDC4811 knockout plants were less resistant to freezing treatment than wild type after acclimation. This suggested that PpCDC481I was an essential gene for low-temperature-induced freezing tolerance in P. patens cells.
基金supported by the National Basic Research Program of China(Grant No.2007CB935700)
文摘In this work,we parallelly detected the specific binding between microarray targets including 12 different kinds of proteins and the probe solution containing five corresponding antibodies and quantitatively analyzed the interactions between CDH13 and solution phase anti-CDH13 at six different probe concentrations by oblique-incidence reflectivity difference(OIRD)method in label-free format.The detection sensitivity reached 10 ng/mL.The experimental results indicate that the OIRD method is a promising and competing technique not only in research work but also in clinic.
