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Syfilis är en sexuellt överförbar infektion som orsakas av spiroketen Treponema pallidum subspecies pallidum. Laboratoriediagnostik utgörs i första hand av serologiska test tillsammans med kliniska fynd men på grund av olika faktorer är dessa inte helt pålitliga. Ett flertal olika PCR-metoder utvecklats som påvisar patogenen med hjälp av T-pallidum-specifika gener. En väl studerad gen med hög specificitet är polA-genen som anses vara en mycket robust och känslig metod och lämpar sig väl för att påvisa T. pallidum i kliniska material. Syftet med projektet är att utveckla och optimera en kvalitativ realtids-PCR i singelplex format för verifiering av T. pallidum, för diagnostisering av syfilis. I detta projekt har en realtids-PCR riktad mot polA för detektion av T. pallidum utvecklats. Metodens prestanda utvärderades med en kommersiellt framställd DNA-kontroll med avseende på sensitivitet, specificitet detektions-gräns. När samtliga moment verifierats och fastställts testades metoden på kliniskt material från sår. Den T. pallidum-specifika PCR-analysen visade att det inte förekom korsreaktivitet mot andra agens som förväntas finnas i sår från patient med misstänkt syfilis. Metoden uppvisade en känslighet vid spädning 1:100 (cirka 13 kopior/µl) och en precision på 36,3±0,8 Ct. Det kliniska provet visade på förekomst av T. pallidum dock är känsligheten något sämre än existerande referensmetod. Metoden kan användas i rutindiagnostik av T. pallidum dock bör utfallet från extraktion av DNA från sår studeras ytterligare för att eventuellt öka utbytet och detektera lägre koncentrationer i kliniskt material. / Syphilis is a sexually transmitted infection caused by the spirochete Treponema pallidum subsp. pallidum. Laboratory diagnostics consist primarily of serological tests together with clinical findings for different reasons these methods are not reliable. A variety of PCR methods has been developed that target the pathogen using T. pallidum-specific genes. A well-studied gene with high specificity is the polA gene which is considered very robust and sensitive method and well suited for detection of T. pallidum in clinical materials. The aim of the project is to develop and optimize a qualitative real-time PCR in single-plex format for the verification of T. pallidum, for the diagnosis of syphilis. A real-time PCR targeting polA was developed for detection of T. pallidum. An evaluation of the method's performance was done with a commercially produced DNA control with regards to sensitivity, specificity detection limit and precision. When all test where verified and established, the method was tested on clinical material from ulcers. The results of the T. pallidum-specific PCR-assay showed that there was no cross-reactivity to other agents that are expected to be in ulcers from patients with suspected syphilis. The method showed a sensitivity at dilution 1:100 (about 13 copies/μl) and a precision of 36.3 ± 0.8 Ct.The clinical specimen showed presence of T. pallidum, however, the sensitivity was not as good than the existing reference method. The method can be used in routine diagnostics of T. pallidum, however, the outcome of extraction of DNA from ulcers should be further studied in order to increase the yield and detect lower concentrations in clinical material.

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Abstract Introduction: H.J.-Haase developed a systematic set of tests for evaluating binocular vision called the Pola Test. The Pola Test measures associated phoria and stereoacuity at distance and near using a variety of different targets for each. This testing method and interpretation is referred to as MKH-Haase (Measuring and Correcting Methodology after H.J.Haase ???the MKH) method. The MKH method is more commonly used in Germany and other European countries than English speaking countries. The MKH-Haase method has been considered a reliable method for prescribing prisms to symptomatic binocular vision patients. Purpose: To investigate the test-retest reliability of binocular vision measurements using the MKH-Haase series of tests that comprise the Pola Test. In addition, I will compare the Pola results with other associated phoria and stereoacuity tests used in North America. Methods: Thirty-four symptomatic and 40 asymptomatic subjects (based on a symptoms questionnaire) participated in this study. Associated phoria and stereoacuity with different tests, including the Pola Test at distance and near, were measured for those subjects on two different sessions. Not all of subjects were tested with all tests. Only 30 subjects in each group completed all of tests. The Pola Test protocol requires the associated phoria and stereoacuity to be measured twice within a session; once with the Polariods oriented with their axes at 45o and 135o and again with the axes switched. Results: Within and between-sessions repeatability of MKH-Haase associated phoria and stereoacuity tests results revealed that most of MKH-Haase associated phoria and stereoacuity tests showed good repeatability within and between-sessions at both distance and near. However, there were a few exceptions to this general finding. Distance horizontal associated phoria values for the Cross Test and Pointer Test at the first session, and the distance Double Pointer Test values at the second session showed some differences between the two views. Between-sessions repeatability of the associated phoria tests did not show any significant differences. For the stereoacuity tests, the differences between the two disparities were statistically significant at the first session for the symptomatic group Line Test and asymptomatic group Step Test. For the second session at distance, the differences were significant with Step Test for both groups. The differences between sessions for both disparities were not significant for most of tests. The symptomatic group???s Step Test for crossed disparity and asymptomatic group's Step Test for uncrossed disparity were exceptions. A repeated measures ANOVA test was conducted to compare different associated phoria tests. Horizontal associated phoria tests without central fusion lock were significantly different from those with central fusion lock at distance and near. Comparison of different stereoacuity tests was conducted by comparing the number of subjects who could identify specific stereothreshold values. Results showed that at both distance and near, there were no significant differences between contour and global stereoacuity tests based on number of subjects who could attain 60 sec of arc or better. Discussion and Conclusion: Most of MKH-Haase associated phoria and stereoacuity charts have reasonable within and between-sessions repeatability. However, some associated phoria tests showed some differences especially with subjects who had higher values. Although there was a significant difference between various horizontal associated phoria tests at distance and near, most of the values differed by around 0.50 ???. The exception was the difference between the Wesson Card and Disparometer. The Wesson card was more exo by 1.50 ??? than the Disparometer. Vertical associated phoria tests did not show any significant differences. Although MKH-Haase chart can measure local stereothreshold down to 10 sec of arc at distance, the AO Slide is easier to perceive. Random dot stereoacuity can be measured with MKH-Haase charts at distance down to 30 sec of arc. All of the contour stereoacuity tests are comparable at near. However, the MKH-Haase chart was easier to perceive. The Random Dot Randot test would be more useful for fast screening purposes. Random dot MKH-Haase test would be easier than TNO Test to measure random dot stereothreshold at near.

This thesis is devoted to the rare decay π0 → e+ e− . We firstly introduce all theoretical knowledge necessary for the description of the vertices used in this work. Subsequently, we introduce three two-loop chiral Feynman diagrams, which are of order O(α2 p4 ). These diagrams are firstly decomposed to Master integrals by Laporta algorithm. After that, we calculate each Master integral individually by the differential equation technique. We also focus on the renormalization of the process. Since we are working with the effective field theory, finite corrections χ from counterterm diagrams naturally appear. With use of the calculations listed in this thesis, the determination of the parameter χR could be further improved taking into account the whole two-loop correction. 1