Rapid Capture System

An automated robotic platform for HC, called the Rapid Capture System (RCS), is being developed for high-volume laboratory testing. The Digene Rapid Capture System (RCS) is a robotic, 96-well microplate processor that integrates liquid and plate handling, incubations, shaking, and washing directly from bar-coded primary tubes (Figure 9.11).

Bulk denaturation of specimens is performed directly in the specimen collection tubes, utilizing a custom rack assembly, a multitube rack vortexer, and a 65°C water bath. Following this denaturation step, specimens are then placed into the RCS platform. Processed plates are transferred to the DML 2000tm luminometer for detection and analysis utilizing the Digene qualitative software. The RCS protocol provides over 3.5 h of continuous hands-free time to the user. The automated application allows a single user with one RCS to process up to 352 specimens (four microplates) in an 8-h shift and 704 specimens in a 13-h period. Currently, the HPV, CT, and GC assays have been adapted to this format, and plans are being made to automate HBV testing as well. Results for the CT/GC assays in several independent laboratories are good, and evaluations are proceeding. The RCS method for HC2 HPV testing has produced good "in-house" results on clinical specimens (Figure 9.12), but to date independent performance data are not available.

RCS provides for "walk-away" automation with bar-coded primary tube sampling (post-denatured specimen rack loading), integration of liquid and microplate handling, incubation, plate shaking, and plate washing. The specimens are bar-coded by specialized software for identification and then processed in the robotic platform. Currently, all processing steps occur on the platform, except for the specimen denaturation and plate reading steps.

Digene Rapid Capture
FIGURE 9.11 Rapid Capture System (RCS).

99.2% Agreement

Rapid Capture System Rcs
FIGURE 9.12 Scatterplot comparison of the manual HC2 and the automated RCS for HPV detection from a split panel of identical clinical specimens. The diagonal line passing through the origin shows excellent agreement between the two tests.

9.6 conclusion

In conclusion, HC has evolved into a flexible and accurate method for the routine detection of viruses, bacteria, and genetic changes and for gene expression profiles. An HC robotic platform, the RCS, has been developed to allow automated high-volume laboratory testing. Invention of the HC3 technology has extended the useful range of applications to the detection and discrimination of closely related nucleic acid targets. HC-EAS allows for sensitive and rapid quantification of mRNA for expression profiling. HC is poised to be among the leading technologies used for disease detection at the molecular level for the next decade.

acknowledgments

We thank Iwona Mielzynska-Lohnas and James Lazar for the use of unpublished data and Katherine Mack for assistance in preparation of the manuscript.

references

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