Fully Automated Molecular Diagnostic System "Simprova" for Simultaneous Testing of Multiple Items.

Nucleic acid amplification-based diagnostics is known as one of the molecular diagnostic systems that allows higher sensitive detection of pathogens than test methods such as immunoassay. However, it has not been widely used because it is complicated to use and takes a long time to generate results. On the other hand, development of fully automated molecular diagnostic systems has been growing around the world as demand for such systems from physicians and laboratory technicians has increased. To meet this demand, we have developed the "Simprova" fully automated molecular diagnostic system, which takes advantage of LAMP (Loop-mediated Isothermal Amplification), a method Eiken Chemical Co., Ltd. invented. Simprova comprises a master unit that controls the entire system and a test unit that extracts and purifies nucleic acid from samples (pretreatment), and uses the LAMP method to detect and amplify nucleic acid. Users can obtain test results automatically by simply installing a pretreatment cartridge, a multi-well testing chip and the sample in the test unit. The multi-well testing chip has 25 reaction wells connected by channels and enables simultaneous testing of multiple targets with one sample. Turnaround time for one test is approximately 30 minutes. Since a conventional extraction and purification method using magnetic-bead separation is used for the pretreatment, nucleic acid can be extracted from serum, plasma, whole blood, urine, and sputum, for example. In addition, the system can perform random-access testing by connecting four test units to the master unit to realize near-the-patient testing. Simprova is therefore a robust and useful system for a wide variety of applications.

Publisher Correction: Clinical evaluation of fully automated molecular diagnostic system "Simprova" for influenza virus, respiratory syncytial virus, and human metapneumovirus.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Clinical evaluation of fully automated molecular diagnostic system "Simprova" for influenza virus, respiratory syncytial virus, and human metapneumovirus.

Influenza virus, respiratory syncytial virus, and human metapneumovirus commonly cause acute upper and lower respiratory tract infections, especially in children and the elderly. Although rapid antigen detection tests for detecting these infections have been introduced recently, these are less sensitive than nucleic acid amplification tests. More recently, highly sensitive point-of-care testings (POCTs) have been developed based on nucleic acid amplification tests, which are easy to use in clinical settings. In this study, loop-mediated isothermal amplification (LAMP)-based POCT "Simprova" to detect influenza A and B viruses, respiratory syncytial virus, and human metapneumovirus was developed. Simprova system is fully automated and does not require skilled personnel. In addition, positive results can be achieved faster than with PCR. In this study, the accuracy of the POCT was retrospectively analyzed using 241 frozen stocked specimens. Additionally, the usability of the Simprova at clinical sites was assessed in a prospective clinical study using 380 clinical specimens and compared to those of real-time PCR and rapid antigen detection test. The novel LAMP-based POCT demonstrated high sensitivity and specificity in characterizing clinical specimens from patients with influenza-like illnesses. The Simprova is a powerful tool for early diagnosis of respiratory viral infections in point-of-care settings.

[Endoscopic endonasal sinus surgery using a navigation system for pituitary adenoma].

Hardy's operation using a microscope has long been the standard for treating pituitary adenoma. A new endonasal approach to the sella using an endoscope combined with a navigation system has been reported, which we used to conduct endocopic endonasal hypophysectomy from October 2000 to June 2003 in 11 patients with pituitary lesions. We introduced an angle-dependent navigation system, Neuro Navigator III. We approached the sphenoid sinus mainly via the hemilateral common meatus. The deviation of the nasal septum and sphenoidal septum was carefully evaluated to determine the optimal operating side. We concluded that the hemilateral common meatus route is useful because it is least invasive in endocopic endonasal hypophysectomy. Another route should be taken, however, if hemorrhaging is uncontrollable or the tumor is quite large. Navigation systems are quite effective in executing this operation safely. Angle-dependent navigation system is a good choice for this operation, considering its cost performance.

Posterior pelvic tilt in patients with decreased lumbar lordosis decreases acetabular femoral head covering.

The relationship between pelvic tilt and acetabular femoral head covering was determined in 38 patients with decreased lumbar lordosis. Posterior pelvic tilt and acetabular femoral head covering were measured on radiographs taken in both supine and standing positions. Previously reported normal values obtained from 96 patients were used as controls. Posterior pelvic tilt increased (P<.01) in patients with decreased lumbar lordosis and anterior acetabular femoral head covering decreased (P<.0001) compared with the controls. These differences were more remarkable in the standing position than in the supine position. Patients with decreased lumbar lordosis have decreased femoral head covering, especially in the standing position.

A new microcolumn-type microchip for examining the expression of chimeric fusion genes using a nucleic acid sandwich hybridization technique.

We report a new type of microcolumn installed in a microchip. The architecture allows use of a nucleic acid sandwich hybridization technique to detect a messenger RNA (mRNA) chain as a target. Data are presented that demonstrate that the expression of a chimeric fusion gene can be detected. The microcolumn was filled with semi-transparent microbeads made of agarose gel that acted as carriers, allowing increased efficiency of the optical detection of fluorescence from the microcolumn. The hybrid between the target trapped on the microbeads and a probe DNA labeled with a fluorescent dye was detected by measuring the intensity of the fluorescence from the microcolumn directly. These results demonstrate an easy and simple method for determining the expression of chimeric fusion genes with no preamplification.

Asymmetric aldol reaction via memory of chirality.

Asymmetric aldol reactions of α-amino acid derivatives via memory of chirality were developed. Chiral oxazolidones with contiguous tetra- and trisubstituted chiral centers were obtained in 78-94% ee by the asymmetric aldol reaction followed by intramolecular acylation.

Point-of-care testing system enabling 30 min detection of influenza genes.

We developed a portable and easy-to-use nucleic acid amplification test (NAT) system for use in point-of-care testing (POCT). The system shows sensitivity that is sufficiently higher than that of the currently available rapid diagnostic kit and is comparable to that of real-time reverse transcription polymerase chain reaction (RT-PCR) for influenza testing.