Rapid detection of mecA and spa by the loop-mediated isothermal amplification (LAMP) method.

AIM: To develop a detection assay for staphylococcal mecA and spa by using loop-mediated isothermal amplification (LAMP) method. METHODS AND RESULTS: Staphylococcus aureus and other related species were subjected to the detection of mecA and spa by both PCR and LAMP methods. The LAMP successfully amplified the genes under isothermal conditions at 64 degrees C within 60 min, and demonstrated identical results with the conventional PCR methods. The detection limits of the LAMP for mecA and spa, by gel electrophoresis, were 10(2) and 10 cells per tube, respectively. The naked-eye inspections were possible with 10(3) and 10 cells for detection of mecA and spa, respectively. The LAMP method was then applied to sputum and dental plaque samples. The LAMP and PCR demonstrated identical results for the plaque samples, although frequency in detection of mecA and spa by the LAMP was relatively lower for the sputum samples when compared to the PCR methods. CONCLUSION: Application of the LAMP enabled a rapid detection assay for mecA and spa. The assay may be applicable to clinical plaque samples. SIGNIFICANCE AND IMPACT OF THE STUDY: The LAMP offers an alternative detection assay for mecA and spa with a great advantage of the rapidity.

Antigenic group II chaperonin in Methanobrevibacter oralis may cross-react with human chaperonin CCT.

Methanobrevibacter oralis is an archaeal species frequently isolated from sites of severe periodontitis. However, its pathogenic roles remain unclear. Here, we aimed to isolate group II chaperonin from M. oralis and examine its antigenicity. The genes encoding two chaperonin subunits (Cpn-1 and Cpn-2) were cloned from M. oralis using polymerase chain reaction and genome walking procedures. Recombinant proteins Cpn-1 and Cpn-2 were generated, and the reactivities of sera from patients with periodontitis were examined by Western immunoblotting. The open reading frames of Cpn-1 and Cpn-2 genes consisted of 1641 and 1614 base pairs, respectively. Putative ATP-binding domains conserved among the chaperonin family were observed in both genes. The deduced amino acid sequences of the two genes showed 28.8-40.0% identity to each of the subunits of human CCT (CCT1-8). Thirty and 29 of 36 patients' sera reacted with the recombinant Cpn-1 and recombinant Cpn-2, respectively. Western immunoblotting using antiserum against human CCT subunits indicated that anti-CCT3 and anti-CCT8 antibodies recognized recombinant Cpn-1. In addition, anti-CCT1, CCT3, CCT6, and CCT8 antibodies recognized an antigen of approximately 60 kDa in M. oralis. The results suggested that the chaperonin subunits of M. oralis were antigenic molecules that were recognized by periodontitis patients and that may cross-react with human chaperonin CCT.

[On eruption of mandibular third molar after extraction of mandibular first or second molar].

In the orthodontic practice, the first or second molars are extracted by reason of a necessity based on the treatment planning. In these cases, it is desirable that the third molar would be taken part in the masticatory function. This study was made to investigate the natural movement of the erupting third molars after the extraction of the first or second molar on the serial lateral cephalometric radiographs and orthopantomographs. Subjects were divided into three groups. The first is the group which the mandibular second molars were extracted. The second is the group which the first molars were extracted bringing about the successful eruption of the third molars. The third is the group which the first molars were extracted with resultant in the failure of the third molar eruption. In all cases, the orthodontic forces were not applied to the third molars. The results were as follows: 1. In the first group, all third molars were successfully erupted and a lot of the spaces after the extraction of second molars were utilized for the eruption of third molars. 2. In the second group, all third molars were successfully erupted and a lot of the spaces after the extraction of first molars were utilized for the mesial movement of the second molars. 3. In the third group, there were various patterns of the impaction of the third molars. It was suggested that the eruption of the third molar was related to the space distal to the second molar but it was not related to the anterio-posterior length of the mandibular body significantly.

[Recognition of orthodontic patients and their parents about the orthodontic treatment and results--a questionnaire method].

This study was undertaken to investigate the desire and consciousness of orthodontic patients and their parents on the content and effect of orthodontic treatment. The subjects were 362 post-treatment patients and 353 of their parents, who answered the questionnaires. The following conclusions were obtained: 1. 49% of the patients and 16% of the parents of the patients thought of giving up the treatment while the patients were under the orthodontic treatment. The main reasons were the discomfort of orthodontic appliances, long treatment period, and the absence from school. 2. 55% of the patients felt uneasy about a change of the occlusion for the worse after removal of the orthodontic appliances, and 60-70% of the patients and parents had forgotten the necessity of the retainer after active orthodontic treatment. 3. Both patients and parents hoped to finish the orthodontic treatment by the end of junior high school. 4. About 50% of the parents preferred the university hospital and about 45% preferred a private dental office which was convenient for attending as an outpatient. 5. About 70% of the patients and parents were satisfied with occlusion after orthodontic treatment, and 33% of the patients and 61% of the parents were satisfied with the orofacial appearance after treatment. 6. About 9% of the patients hoped to keep secret their history of orthodontic treatment. 7. About 35% of the patients and 60% of the parents would recommend people with malocclusions to receive orthodontic treatment. About 80% of the patients would make their own children receive the orthodontic treatment if needed. 8. About 90-95% of the patients and parents were pleased with orthodontic treatment.