TRACING THE CRIMINAL Part nine: Criminals in spiral form Institute for microbiology shows From anthem of medical students „Diabetes mellitus, icterus et vomitus“ (Second part) Treponema pallidum Gonococcus ruber Ulcus molle, ulcus durum Molle est reparaturum Nos curabit … (name of a suitable urologist of dermatovenerologist) Survey of topics Clinical characteristics of spiral bacteria Story one n Roseanne Pinkspot started to have pink spots on her body. She thought, that probably… Oh yes, several weeks ago whe participated on a girl scout camp and several times during the camp she had a tick. n Her GP sent her to children infection clinic, and experienced infectionist confirmed, that most likelly it is the disease that Roseanne supposed. For sure, she took serum for antibody detection… Erythema migrans n This is a picture of Erythema migrans of student M. M., who kindly agreed to let it for use in education The causative agent was Lyme disease – tick borne zoonosis Story two (virtual, but basis is from a real story) n When Phyllis found, that she really needs pervitin, and more and more, she decided to earn money by her own body. n When the client paid more, she went with him without a preservative, she used anticonception and she felt more OK n Then she fell in love and decided to have a child. She stopped the anticonception and was happy. Helmut will be a good father… Story two – continuing n So Phyllis was pregnant. But she found heerself a genital ulcus and her gynekologist took blood for serological examination. It was positive. Phyllis did not want interruption, it was too late and she wanted her child. n Phyllis was treated, but the antibiotic was wrong. The child was born ill and after two weeks it died because of a secondary Klebsiella septicaemia The criminal was n Treponema pallidum, causing syfilis n Syfilis is a classic sexual disease. It is transmitted sexually only. But it is a systemic disease – in developped stages the whole body is affected (gummata, aortal dissection, neurolues, psychical symptomas) n Some subspecies of T. pallidum and some other treponemas cause other diseases (framboesia – yaws, T. pertenue) n Some treponemas are non-pathogenous Course of syphilis Course of syphilis Tertiary syphilis Lues Yaws (framboesia) Story three n Mr. Ratter was an employee of NWPS Ltd. (Nowhere Water Pipes and Sewage) n His job was sewage cleaning. He knew all sewage corridors. He also knew rat habits, he liked rats and he understood them. n Nevertheless, once there was some mis-understanding between him and the leader of rat group and Mr. Ratter was bitten to his leg. n Some time after this, Mr. Ratter was hospitalized with icterus and bleeding… This is not Mr. Ratter, but his Venezuelan colleague with a similar fate… The disease is caused by… Microbiologic characteristics and diagnostics of spirochets Spirochets Treponema: direct methods n Direct diagnostics is rare, also because often there is hardly something to take. Only patients with chancre are available fot scrapping. n Microscopy: It is possible to use wet mount – dark field. It is strange, that although is is a wet mount, immersion is used (treponemas are very subtle). Besides that, fluorescence staining can be used n Neither culture nor biochemical methods are used n Antigen detection can be performed by direct IMF n Animal experiment: There exist so named RIT – Rabbit infectivity test n PCR diagnostics is more and more important Direct diagnostics of syphilis n RIT – Rabbit infectivity test. For ethical reasons, but also as it is too much work, the RIT is minimized today. n Dark field – shining Treponema pallidum is s observed againts the dark field n Direct IMF – another direct, but difficult method New Zealand Rabbit used for RIT Dark field microscopy Immunofluorescence Treponema: indirect methods n We use non-treponema tests, where cardiolipin from bovine hearths acts as an antigen, and treponema tests, where we have a real antigen from Treponema pallidum n Diagnostics is composed of screening and confirmation. We confirm everything that was positive or at least borderline at screening, in reasonable cases even negative results. n Screening usually consists of a non-treponema and a treponema test n Confirmation is performed by highly specific treponema tests The most important indirect tests for lues RRR and TPHA n In RRR, the well with turbidity is positive (it looks like the positive control). It is necessary shake the panel, otherwise the reaction would not be visible! n TPHA is an agglutination on carrier (RBC). A „potato shaped formation“ is positive, a dense dot is negative TPHA – to remember To remember TPHA II (www.medmicro.info) TPHA – reading: Indications for confirmation n Screening reactions are performed allways, when somebody is to be tested for syphilis (including e. g. pregnant women that are not at all supposed to be positive). Screening reactions are performed only qualitatively or semiquantitatively n Indication for confirmation is: n any positive or at least borderlinde result in RRR and/or TPHA reaction n presence of suspicious lesions on body, or anamnesis of risky sexual intercourse – here even in case of negativity of both reactions ELISA, Western blotting and PCR in spirochetal diagnostics n ELISA, Western blotting and PCR – all of them are used in spirochets simillarly as in other cases – see J10 and J11 topics in spring term. n Positive are patients with values of absorbance higher than a given value (calibration well, cut off etc.) n Examination of IgG and IgM antibodies is important, mere IgG positivity is just a proof of a previous infection. n PCR is used in diagnostics of syphilis and Lyme disease. It is usually positive sooner than methods detecting antibodies. Borrelia and leptospira – course of investigation n Borrelia: Mostly serology, event. PCR. In serology, IgM (typical fot an early infection) and IgG antibodies are detected using ELISA method, positive finding is confirmed by Western blotting. Western blotting is more specific. n Leptospira: Dark field microscopy and culture in special medium are used Leptospiral diagnostics n Microscopy of leptospira More diagnostic opportunities in leptospira (latex agglutination) Pictures of spiral bacteria Galery of criminals Borrelia burgdorferi Borrelia recurrentis Treponema pallidum Leptospira interrogans Leptospira interrogans Leptospira interrogans Leptospira in the electronoptic microscope The End