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اهلاً وسهلاً بالحضور الكرام. اهلاوسهلا بالحضور الكرام. Welcome. Dr.Sundus Nsaif AL- Hucheimi College of Medicine University of Kufa. بسم الله الرحمن الرحيم س ُبحَانَكَ لا عِلمَ لَنا إِلا ماَ عَلَمَّتناَ إِّنكَ أنت اْلعِلَيِم ألحكيِمَ صدق الله العلي العظيم ) ).

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slide1

اهلاً وسهلاً بالحضور الكرام

اهلاوسهلا بالحضور الكرام

Welcome

slide2

Dr.SundusNsaifAL-Hucheimi

College of Medicine

University of Kufa

slide3

بسم الله الرحمن الرحيم

سُبحَانَكَ لا عِلمَ لَنا إِلا ماَ عَلَمَّتناَ إِّنكَ أنت اْلعِلَيِم ألحكيِمَ

صدق الله العلي العظيم))

slide4

Tracking of Cutaneous Leishmaniasis

by Parasitological, Molecular and Biochemical Analysis

slide5

Introduction

Leishmaniasis is one of the infectious parasitic diseases transmitted by biting sand flies with highest incidence in the world.WHOhas ranked leishmaniasis as one of the six important infectious diseases .

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There are an estimated number of 12 million cases worldwide, with 2 million new cases added each year .90% of all cases occur in Afghanistan, Brazil, Iran ,Iraq, Saudi Arabia, and Syria .

  • *CL was mostly present in central Iraq and the Greater Baghdad area, but since the Gulf War the disease has extended to new areas rarely affected before, such as Missan, Thi-Qar, and Basrah .
  • *The number of CL cases doubled in study area after 2004,and caused several outbreak such as epidemic outbreaks have been reported in Qadessia province in 2008 .
  • *Leishmaniasis still constitute a major public health problem and underreported to a large degree.
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Aims of study

The present study aims at :

1.The specification of the different species and strain of genus Leishmania by nested –PCR and cellulose acetate electrophoresis.

2.The evaluation of the incidence and epidemiological aspects of cutaneous leishmaniasis in the study area .

3.The detection and containing epidemics in the early stages and providing early diagnosis .

slide9

The study included 126 patients with lesions clinically suggestive to be cutaneous leishmaniasis whom attended to hospitals from five Iraqi provinces as following: Al-Najaf , Babylon , Al-Qadisya, Karbala and Kut province .

parasitology

Culture

NNN,Schneider

Drosphila,RBMI1640

Direct smear examination

positive

positive

negative

negative

biochemical

molecular

slide10

Culture

NNN,SchneiderDrosphila ,RBMI1640

Molecular test

biochemical test

DNA EXTRACTION

by using the Bioneer Genomic DNA extraction Kit

The comparisons were made by examination of the CAE of 4 soluble enzymes

Nested-pcr

LP

6PGDH

GPI

First step to detect leishmania genus

Second step to identify leishmania spp.

MPI

slide12

Distribution of the type of infection according

to type of lesions and age groups ofpatients.

* S = Single , M = Multiple

slide14

Direct smear examination

according to sex and age groups ofpatients.

slide15

Direct skin smear (Giemsa stain , 100 x)

  • showing amastigotesin lesion aspiration
slide17

Direct skin smear (Giemsa stain , 100 x) showing amastigotesin discharged of blood

slide21

Nested-PCR (first step)

according to sex and age groups ofpatients.

slide22

Nested-PCR (second step)

according to sex and age groups ofpatients.

slide23

Agarose gel electrophoresis ofLeishmaniaisolates in nested-PCR(first step) .Lane 1-15Leishmaniaspp , Lane 16,DNA size marker 100Pb

slide24

Agarose gel electrophoresis of Leishmania isolates in nested-PCR(second step) .Lane 1,13 ,DNA size marker 100 Pb;Lane 2,negative control ;Lane 3,6,7,11 (560Pb) L.majorisolates;Lane 4,5,8,9,10,12(750Pb)L. tropicaisolates

slide25

Cellulose acetate electrophoresis

according to sex and age groups ofpatients.

slide26

Conclusion

Nested-PCR is a reliable method for the diagnosis and identification of Leishmania species and can be applied in epidemiologic investigations.

The cellulose acetate electrophoresis (isoenzymeanalysis) was an ideal method for discrimination of Leishmania. spp. variants.

slide27

Conclusion

3. Both L. major and L. tropicawere the causative agents of cutaneous leishmaniasis in study area .

4.L.major was the main species causing CL in comparing with L.tropica in the present study.

slide28

Recommendations

1. Opening number of research centres situated in the country offer DNA applications for so-called “parasite tracking.” After collection in the field, samples can be simply transferred to such a centre for further analysis.

slide29

Recommendations

2. More experimental studies for effective vaccine are developed for leishmaniasis.

slide30

Recommendations

3. Control of sand flies through residual insecticides spraying, Since L. major(the dominant species.) is an zoonotic parasite, therefore, it is emphasize on rodents eradication program and treatment of patients.

slide31

Recommendations

4. General physicians must be have a clear idea about diagnosis and treatment besides a fair knowledge about its pathophysiology specially in rural areas where this disease is most prevalent.

slide32

Recommendations

5. To understand the atypical behavior of Leishmania spp. in Iraq, studies need to be directed toward understanding the vector bionomics and reservoir hosts for this parasite.