Dataset Information

Quantification of SSX mRNA expression in human bone and soft tissue tumors using nucleic acid sequence-based amplification.

ABSTRACT: The SSX family proteins have been considered new members of the cancer/testis antigens because of the restricted expression in testis among normal tissues and the activation in a wide range of cancers. Thus, they would be potential molecular targets for immunotherapeutic strategies. We have developed a competitive nucleic acid sequence-based amplification (NASBA) assay to analyze SSX mRNA expression in 211 bone and soft tissue tumors. The copy numbers of SSX mRNA per mug of total RNA in tumor tissues were widely distributed, ranging logarithmically from 0.6 to 6.6. We found that malignant tumors showed significantly higher expression of SSX mRNA than benign tumors (P < 0.0001). Further, SSX mRNA expression in stage III tumors was significantly higher than that in stage I or II tumors (P < 0.005). This NASBA assay was also more sensitive compared to immunohistochemistry using newly affinity-purified polyclonal antibody against SSX. Collectively, these results suggest that the SSX quantitative NASBA assay could provide useful information to select eligible patients for SSX-specific cancer vaccines.

SUBMITTER: Naka N

PROVIDER: S-EPMC1867521 | biostudies-literature | 2005 May

REPOSITORIES: biostudies-literature

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Quantification of SSX mRNA expression in human bone and soft tissue tumors using nucleic acid sequence-based amplification.

Naka Norifumi N Joyama Susumu S Tsukamoto Yoshitane Y Yoshioka Kiyoko K Hashimoto Nobuyuki N Ujiiye Takeshi T Hayashi Tsukasa T Kawase Masako M Mano Masayuki M Ishiguro Shingo S Myoui Akira A Ueda Takafumi T Yoshikawa Hideki H Araki Nobuhito N Itoh Kazuyuki K

The Journal of molecular diagnostics : JMD 20050501 2

The SSX family proteins have been considered new members of the cancer/testis antigens because of the restricted expression in testis among normal tissues and the activation in a wide range of cancers. Thus, they would be potential molecular targets for immunotherapeutic strategies. We have developed a competitive nucleic acid sequence-based amplification (NASBA) assay to analyze SSX mRNA expression in 211 bone and soft tissue tumors. The copy numbers of SSX mRNA per mug of total RNA in tumor ti ...[more]

PMID: 15858142

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Project description:Bacterial infection is a widespread problem in humans that can potentially lead to hospitalization and morbidity. The largest obstacle for physicians/clinicians is the time delay in accurately identifying infectious bacteria, especially their sub-species, in order to adequately treat and diagnose such infected patients. Loop-mediated amplification (LAMP) is a nucleic acid amplification method that has been widely used in diagnostic applications due to its simplicity of constant temperature, use of up to 4 to 6 primers (rendering it highly specific), and capability of amplifying low copies of target sequences. Use of interfacial effect-based monitoring is expected to dramatically shorten the time-to-results of nucleic acid amplification techniques. In this work, we developed a LAMP-based point-of-care platform for detection of bacterial infection, utilizing smartphone measurement of contact angle from oil-immersed droplet LAMP reactions. Whole bacteria (Escherichia coli O157:H7) were assayed in buffer as well as 5% diluted human whole blood. Monitoring of droplet LAMP reactions was demonstrated in a three-compartment, isothermal proportional-integrated-derived (PID)-controlled chip. Smartphone-captured images of droplet LAMP reactions, and their contact angles, were evaluated. Contact angle decreased substantially upon target amplification in both buffer and whole blood samples. In comparison, no-target control (NTC) droplets remained stable throughout the 30?min isothermal reactions. These results were explained by the pre-adsorption of plasma proteins to an oil-water interface (lowering contact angle), followed by time-dependent amplicon formation and their preferential adsorption to the plasma protein-occupied oil-water interface. Time-to-results was as fast as 5?min, allowing physicians to quickly make their decision for infected patients. The developed assay demonstrated quantification of bacteria concentration, with a limit-of-detection at 102?CFU/?L for buffer samples, and binary target or no-target identification with a limit-of-detection at 10?CFU/?L for 5% diluted whole blood samples.

Dataset Information

Quantification of SSX mRNA expression in human bone and soft tissue tumors using nucleic acid sequence-based amplification.

Publications

Quantification of SSX mRNA expression in human bone and soft tissue tumors using nucleic acid sequence-based amplification.

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