Molecular detection and genotyping of Toxooplasma gondiistrains isolated in Serbia
Toxoplasmosis, infection caused by the parasite Toxoplasma gondii, is one of the most widespread zoonoses in the world and it has been estimated that up to one third of the global human population is infected with this parasite. Although serological tests are the primary method in the routine diagno...
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| description | Toxoplasmosis, infection caused by the parasite Toxoplasma gondii, is one of
the most widespread zoonoses in the world and it has been estimated that up
to one third of the global human population is infected with this parasite.
Although serological tests are the primary method in the routine diagnosis of
toxoplasmosis, interpretation of the results can be not only difficult and
time-consuming, but they can be inconclusive. Therefore, the introduction of
highly sensitive molecular methods, by which results can be obtained within a
day, into the diagnosis of toxoplasmosis is of great importance. The aim of
this research was the introduction of molecular methods into the diagnosis of
toxoplasmosis for the first time in Serbia, as well as genotypization of the
isolated T. gondii strains. This study included 160 human biological samples
from 127 patients serologically suspected of toxoplasmosis in which the
presence of T. gondii was analyzed by PCR and by bioassay in mice. It was
found that the detection of T. gondii DNA in human biological materials
depends on the type of sample as well as on the initial volume. Real-time PCR
was positive in 30.8% of peripheral blood samples, 33.3% of cord blood
samples, 35.7% of amniotic fluid samples, 57.1% of CSF samples and 60% of
humor aqueous samples. The detection rate of T. gondii by molecular methods
was higher than by bioassay, in both blood samples from adults suspected of
acute toxoplasmosis and samples from fetuses and newborns suspected of
congenital toxoplasmosis. The results of this study emphasize the huge
practical importance and potential of molecular diagnostics and show the
benefit of introducing this method as part of the standard protocol for the
diagnosis of toxoplasmosis. T. gondii has a wide range of host organisms,
including all warm-blooded animals, so that in the second part of this
research molecular methods for the detection of this parasite were applied in
animal material, rodents specifically. Rodents were chosen because of their
potentially important role as a link in the transmission chain of T. gondii
infection. A total of 156 brain samples from 144 brown rats (Rattus
norvegicus) and 12 house mice (Mus musculus) were analyzed. T. gondii DNA was
vi detected in 10.4% of rats and in as many as 83.3% of house mice. The rate
of positive findings in rats and mice collected in Belgrade is an indicator
of the degree of contamination of the urban environment with T. gondii,
reflecting a considerable |
| format | Dissertation |
| fullrecord | <record><control><sourceid>europeana_1GC</sourceid><recordid>TN_cdi_europeana_collections_9200447_BibliographicResource_3000095543002</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>9200447_BibliographicResource_3000095543002</sourcerecordid><originalsourceid>FETCH-europeana_collections_9200447_BibliographicResource_30000955430023</originalsourceid><addsrcrecordid>eNqtjU0KwjAQRrtxIeod5gJC6Q_iVlG6cVO7D9NkGgdiJiQp6O0t6BH8Nu-t3rcu-ps40rPDCIYy6cziAb0BS17yO7C3IBMM8hIJDtMTwYo3zClHZJ-AkzjMZIA93CmOjNtiNaFLtPtxU3TXy3Du9jRHCYQelRbnvl9JHauybJqDOvHoWGzE8GDdU5I5alJ1uezYts0iVf3H1AcLa1TL</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>dissertation</recordtype></control><display><type>dissertation</type><title>Molecular detection and genotyping of Toxooplasma gondiistrains isolated in Serbia</title><source>Europeana Collections</source><creator>Vujanić Marija</creator><creatorcontrib>Vujanić Marija ; Ivović Vladimir ; Savić-Pavićević Dušanka ; Đurković-Đaković Olgica</creatorcontrib><description>Toxoplasmosis, infection caused by the parasite Toxoplasma gondii, is one of
the most widespread zoonoses in the world and it has been estimated that up
to one third of the global human population is infected with this parasite.
Although serological tests are the primary method in the routine diagnosis of
toxoplasmosis, interpretation of the results can be not only difficult and
time-consuming, but they can be inconclusive. Therefore, the introduction of
highly sensitive molecular methods, by which results can be obtained within a
day, into the diagnosis of toxoplasmosis is of great importance. The aim of
this research was the introduction of molecular methods into the diagnosis of
toxoplasmosis for the first time in Serbia, as well as genotypization of the
isolated T. gondii strains. This study included 160 human biological samples
from 127 patients serologically suspected of toxoplasmosis in which the
presence of T. gondii was analyzed by PCR and by bioassay in mice. It was
found that the detection of T. gondii DNA in human biological materials
depends on the type of sample as well as on the initial volume. Real-time PCR
was positive in 30.8% of peripheral blood samples, 33.3% of cord blood
samples, 35.7% of amniotic fluid samples, 57.1% of CSF samples and 60% of
humor aqueous samples. The detection rate of T. gondii by molecular methods
was higher than by bioassay, in both blood samples from adults suspected of
acute toxoplasmosis and samples from fetuses and newborns suspected of
congenital toxoplasmosis. The results of this study emphasize the huge
practical importance and potential of molecular diagnostics and show the
benefit of introducing this method as part of the standard protocol for the
diagnosis of toxoplasmosis. T. gondii has a wide range of host organisms,
including all warm-blooded animals, so that in the second part of this
research molecular methods for the detection of this parasite were applied in
animal material, rodents specifically. Rodents were chosen because of their
potentially important role as a link in the transmission chain of T. gondii
infection. A total of 156 brain samples from 144 brown rats (Rattus
norvegicus) and 12 house mice (Mus musculus) were analyzed. T. gondii DNA was
vi detected in 10.4% of rats and in as many as 83.3% of house mice. The rate
of positive findings in rats and mice collected in Belgrade is an indicator
of the degree of contamination of the urban environment with T. gondii,
reflecting a considerable public health risk. Finally, this research provides
the very first and original phylogenetic data on T. gondii population
structure in Serbia. Recent studies performed world-wide indicate the
existence of a clonal population structure characterized by three main,
widespread clonal lineages (I, II and III), of which types I and II are
predominant in Europe and North America. There are also recombinant and
atypical strains that are mainly present in South America and Africa. In this
study, the identification of the T. gondii clonal type was performed in 22
samples, 19 from humans and three from rats, using the PCRRFLP method.
Genotyping was successful in five human samples of which four isolates
originating from cases of congenital toxoplasmosis, toxoplasmosis in
pregnancy and bone marrow transplantation were type II, while one isolate
from a case of congenital toxoplasmosis was type I. These results, first for
the Southeastern Europe, indicate the predominance of genotype II in our
country, as well as in most of the Europe.
Toksoplazmoza, oboljenje koje izaziva parazit Toxoplasma gondii, jedna je od
najrasprostranjenijih zoonoza u svetu, i procenjuje se da je čak jedna
trećina čovečanstva inficirana ovim parazitom. Iako serološki testovi
predstavljaju primarnu metodu u rutinskoj dijagnostici toksoplazmoze,
interpretacija rezultata seroloških analiza u nekim kliničkim slučajevima
može biti komplikovana i dugotrajna, a neretko i nedovoljno informativna.
Zbog toga je uvođenje molekularnih metoda koje inače odlikuje velika
osetljivost i pomoću kojih je moguće dobiti rezultate u kratkom vremenskom
periodu, od velikog značaja za dijagnostiku toksoplazmoze. Otuda je cilj ovog
istraživanja bio uvođenje molekularne dijagnostike toksoplazmoze po prvi put
u Srbiji, kao i genotipizacija izolovanih sojeva T. gondii. Ovom studijom je
obuhvaćeno 160 uzoraka poreklom od 127 pacijenata serološki suspektnih na
toksoplazmozu, kod kojih je prisustvo T. gondii analizirano PCR-om u realnom
vremenu i biološkim ogledom. Pokazano je da uspešnost dokazivanja DNK T.
gondii u materijalu humanog porekla zavisi kako od tipa materijala tako i od
ukupne zapremine materijala od kojeg se uzima uzorak. Tako je PCR u realnom
vremenu bio pozitivan u 30,8% uzoraka periferne krvi, u 33,3% uzoraka krvi iz
pupčanika, u 35,7% uzoraka plodovih voda, u 57,1% uzoraka likvora i u 60%
uzoraka očnih vodica. U odnosu na izolaciju parazita biološkim ogledom,
pokazana je znatno viša stopa detekcije DNK T. gondii, i to i u uzorcima krvi
od pacijenata suspektnih na akutnu toksoplazmozu tako i u onima poreklom od
fetusa i novorođenčadi suspektnih na kongenitalnu toksoplazmozu. Dobijeni
rezultati ukazuju pre svega na veliki potencijal i praktični značaj
molekularne dijagnostike kao i na potrebu uvođenja ove metode kao dela
standardnog protokola za dijagnostiku toksoplazmoze. T. gondii ima veoma
širok spektar domaćina koji uključuje sve toplokrvne životinje, te su u drugom
delu ovog istraživanja molekularne metode za detekciju toksoplazmoze primenjene
na materijalu životinjskog porekla, i to konkretno od glodara. Zbog njihove
potencijalno značajne uloge kao karike u epidemiološkom lancu toksoplazmoze u
okviru ove studije ispitano je i 156 uzoraka poreklom od 144 braon pacova
(Rattus norvegicus) i 12 kućnih miševa (Mus musculus). DNK T. gondii je
detektovana u mozgu 10,4% pacova i kod čak 83,3% kućnih miševa. Relativno
visoka stopa pozitivnih nalaza kod pacova i miševa sakupljenih u Beogradu
ukazuje na kontaminiranost naše sredine parazitom T. gondii koja predstavlja
rizik za javno zdravlje. Poslednji deo ovog rada se odnosi na prva i poptuno
originalna filogenetska istraživanja populacione strukture T. gondii. Naime,
skorašnja istraživanja u svetu ukazuju na postojanje klonske populacione
strukture koju karakterišu tri široko rasprostranjena glavna klonska tipa (I,
II i III), od kojih su tipovi I i II predominantni u Evropi i Severnoj
Americi. Postoje takođe i rekombinantni i atipični sojevi koji su uglavnom
prisutni u Južnoj Americi i Africi. U ovom istraživanju određivanje klonskog
tipa, čime su dobijeni i prvi podaci o populacionoj strukturi T. gondii u
Srbiji, vršeno je korišćenjem PCR-RFLP metode. Analizirano je ukupno 22 uzorka
i to 19 humanog porekla i 3 poreklom od pacova. Uspešno je genotipizirano pet
uzoraka poreklom od ljudi, za koje se pokazalo da četiri pripadaju tipu II a
jedan tipu I. Sojevi tipa II su izolovani iz kliničkih uzoraka bolesnika sa
kongenitalnom toksoplazmozom, toksoplazmozom stečenom u trudnoći i
reaktivacijom infekcije usled post- transplantacione imunosupresije, dok je
soj tipa I izolovan iz jednog uzorka poreklom od kongenitalno inficiranog
novorođenčeta. Ovi rezultati, prvi za čitavo područje Jugoistočne Evrope,
pokazuju da je i u našoj zemlji, kao i u većem delu Evrope, predominantan pre
svega genotip II.</description><language>srp</language><publisher>University of Belgrade, Faculty of Biology</publisher><subject>diagnostics ; dijagnostika ; genotipizacija ; genotyping ; glodari ; human material ; humani materijal ; molecular detection ; molekularna detekcija ; parasite strains ; parazita ; PCR ; PCR-RFLP ; rodents ; sojevi ; toksoplazmoza ; Toxoplasma gondii ; toxoplasmosis</subject><creationdate>2012</creationdate><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://data.europeana.eu/item/9200447/BibliographicResource_3000095543002$$EHTML$$P50$$Geuropeana$$Hfree_for_read</linktohtml><link.rule.ids>311,780,38517,76176</link.rule.ids><linktorsrc>$$Uhttps://data.europeana.eu/item/9200447/BibliographicResource_3000095543002$$EView_record_in_Europeana$$FView_record_in_$$GEuropeana$$Hfree_for_read</linktorsrc></links><search><creatorcontrib>Vujanić Marija</creatorcontrib><title>Molecular detection and genotyping of Toxooplasma gondiistrains isolated in Serbia</title><description>Toxoplasmosis, infection caused by the parasite Toxoplasma gondii, is one of
the most widespread zoonoses in the world and it has been estimated that up
to one third of the global human population is infected with this parasite.
Although serological tests are the primary method in the routine diagnosis of
toxoplasmosis, interpretation of the results can be not only difficult and
time-consuming, but they can be inconclusive. Therefore, the introduction of
highly sensitive molecular methods, by which results can be obtained within a
day, into the diagnosis of toxoplasmosis is of great importance. The aim of
this research was the introduction of molecular methods into the diagnosis of
toxoplasmosis for the first time in Serbia, as well as genotypization of the
isolated T. gondii strains. This study included 160 human biological samples
from 127 patients serologically suspected of toxoplasmosis in which the
presence of T. gondii was analyzed by PCR and by bioassay in mice. It was
found that the detection of T. gondii DNA in human biological materials
depends on the type of sample as well as on the initial volume. Real-time PCR
was positive in 30.8% of peripheral blood samples, 33.3% of cord blood
samples, 35.7% of amniotic fluid samples, 57.1% of CSF samples and 60% of
humor aqueous samples. The detection rate of T. gondii by molecular methods
was higher than by bioassay, in both blood samples from adults suspected of
acute toxoplasmosis and samples from fetuses and newborns suspected of
congenital toxoplasmosis. The results of this study emphasize the huge
practical importance and potential of molecular diagnostics and show the
benefit of introducing this method as part of the standard protocol for the
diagnosis of toxoplasmosis. T. gondii has a wide range of host organisms,
including all warm-blooded animals, so that in the second part of this
research molecular methods for the detection of this parasite were applied in
animal material, rodents specifically. Rodents were chosen because of their
potentially important role as a link in the transmission chain of T. gondii
infection. A total of 156 brain samples from 144 brown rats (Rattus
norvegicus) and 12 house mice (Mus musculus) were analyzed. T. gondii DNA was
vi detected in 10.4% of rats and in as many as 83.3% of house mice. The rate
of positive findings in rats and mice collected in Belgrade is an indicator
of the degree of contamination of the urban environment with T. gondii,
reflecting a considerable public health risk. Finally, this research provides
the very first and original phylogenetic data on T. gondii population
structure in Serbia. Recent studies performed world-wide indicate the
existence of a clonal population structure characterized by three main,
widespread clonal lineages (I, II and III), of which types I and II are
predominant in Europe and North America. There are also recombinant and
atypical strains that are mainly present in South America and Africa. In this
study, the identification of the T. gondii clonal type was performed in 22
samples, 19 from humans and three from rats, using the PCRRFLP method.
Genotyping was successful in five human samples of which four isolates
originating from cases of congenital toxoplasmosis, toxoplasmosis in
pregnancy and bone marrow transplantation were type II, while one isolate
from a case of congenital toxoplasmosis was type I. These results, first for
the Southeastern Europe, indicate the predominance of genotype II in our
country, as well as in most of the Europe.
Toksoplazmoza, oboljenje koje izaziva parazit Toxoplasma gondii, jedna je od
najrasprostranjenijih zoonoza u svetu, i procenjuje se da je čak jedna
trećina čovečanstva inficirana ovim parazitom. Iako serološki testovi
predstavljaju primarnu metodu u rutinskoj dijagnostici toksoplazmoze,
interpretacija rezultata seroloških analiza u nekim kliničkim slučajevima
može biti komplikovana i dugotrajna, a neretko i nedovoljno informativna.
Zbog toga je uvođenje molekularnih metoda koje inače odlikuje velika
osetljivost i pomoću kojih je moguće dobiti rezultate u kratkom vremenskom
periodu, od velikog značaja za dijagnostiku toksoplazmoze. Otuda je cilj ovog
istraživanja bio uvođenje molekularne dijagnostike toksoplazmoze po prvi put
u Srbiji, kao i genotipizacija izolovanih sojeva T. gondii. Ovom studijom je
obuhvaćeno 160 uzoraka poreklom od 127 pacijenata serološki suspektnih na
toksoplazmozu, kod kojih je prisustvo T. gondii analizirano PCR-om u realnom
vremenu i biološkim ogledom. Pokazano je da uspešnost dokazivanja DNK T.
gondii u materijalu humanog porekla zavisi kako od tipa materijala tako i od
ukupne zapremine materijala od kojeg se uzima uzorak. Tako je PCR u realnom
vremenu bio pozitivan u 30,8% uzoraka periferne krvi, u 33,3% uzoraka krvi iz
pupčanika, u 35,7% uzoraka plodovih voda, u 57,1% uzoraka likvora i u 60%
uzoraka očnih vodica. U odnosu na izolaciju parazita biološkim ogledom,
pokazana je znatno viša stopa detekcije DNK T. gondii, i to i u uzorcima krvi
od pacijenata suspektnih na akutnu toksoplazmozu tako i u onima poreklom od
fetusa i novorođenčadi suspektnih na kongenitalnu toksoplazmozu. Dobijeni
rezultati ukazuju pre svega na veliki potencijal i praktični značaj
molekularne dijagnostike kao i na potrebu uvođenja ove metode kao dela
standardnog protokola za dijagnostiku toksoplazmoze. T. gondii ima veoma
širok spektar domaćina koji uključuje sve toplokrvne životinje, te su u drugom
delu ovog istraživanja molekularne metode za detekciju toksoplazmoze primenjene
na materijalu životinjskog porekla, i to konkretno od glodara. Zbog njihove
potencijalno značajne uloge kao karike u epidemiološkom lancu toksoplazmoze u
okviru ove studije ispitano je i 156 uzoraka poreklom od 144 braon pacova
(Rattus norvegicus) i 12 kućnih miševa (Mus musculus). DNK T. gondii je
detektovana u mozgu 10,4% pacova i kod čak 83,3% kućnih miševa. Relativno
visoka stopa pozitivnih nalaza kod pacova i miševa sakupljenih u Beogradu
ukazuje na kontaminiranost naše sredine parazitom T. gondii koja predstavlja
rizik za javno zdravlje. Poslednji deo ovog rada se odnosi na prva i poptuno
originalna filogenetska istraživanja populacione strukture T. gondii. Naime,
skorašnja istraživanja u svetu ukazuju na postojanje klonske populacione
strukture koju karakterišu tri široko rasprostranjena glavna klonska tipa (I,
II i III), od kojih su tipovi I i II predominantni u Evropi i Severnoj
Americi. Postoje takođe i rekombinantni i atipični sojevi koji su uglavnom
prisutni u Južnoj Americi i Africi. U ovom istraživanju određivanje klonskog
tipa, čime su dobijeni i prvi podaci o populacionoj strukturi T. gondii u
Srbiji, vršeno je korišćenjem PCR-RFLP metode. Analizirano je ukupno 22 uzorka
i to 19 humanog porekla i 3 poreklom od pacova. Uspešno je genotipizirano pet
uzoraka poreklom od ljudi, za koje se pokazalo da četiri pripadaju tipu II a
jedan tipu I. Sojevi tipa II su izolovani iz kliničkih uzoraka bolesnika sa
kongenitalnom toksoplazmozom, toksoplazmozom stečenom u trudnoći i
reaktivacijom infekcije usled post- transplantacione imunosupresije, dok je
soj tipa I izolovan iz jednog uzorka poreklom od kongenitalno inficiranog
novorođenčeta. Ovi rezultati, prvi za čitavo područje Jugoistočne Evrope,
pokazuju da je i u našoj zemlji, kao i u većem delu Evrope, predominantan pre
svega genotip II.</description><subject>diagnostics</subject><subject>dijagnostika</subject><subject>genotipizacija</subject><subject>genotyping</subject><subject>glodari</subject><subject>human material</subject><subject>humani materijal</subject><subject>molecular detection</subject><subject>molekularna detekcija</subject><subject>parasite strains</subject><subject>parazita</subject><subject>PCR</subject><subject>PCR-RFLP</subject><subject>rodents</subject><subject>sojevi</subject><subject>toksoplazmoza</subject><subject>Toxoplasma gondii</subject><subject>toxoplasmosis</subject><fulltext>true</fulltext><rsrctype>dissertation</rsrctype><creationdate>2012</creationdate><recordtype>dissertation</recordtype><sourceid>1GC</sourceid><recordid>eNqtjU0KwjAQRrtxIeod5gJC6Q_iVlG6cVO7D9NkGgdiJiQp6O0t6BH8Nu-t3rcu-ps40rPDCIYy6cziAb0BS17yO7C3IBMM8hIJDtMTwYo3zClHZJ-AkzjMZIA93CmOjNtiNaFLtPtxU3TXy3Du9jRHCYQelRbnvl9JHauybJqDOvHoWGzE8GDdU5I5alJ1uezYts0iVf3H1AcLa1TL</recordid><startdate>20120709</startdate><enddate>20120709</enddate><creator>Vujanić Marija</creator><general>University of Belgrade, Faculty of Biology</general><scope>1GC</scope></search><sort><creationdate>20120709</creationdate><title>Molecular detection and genotyping of Toxooplasma gondiistrains isolated in Serbia</title><author>Vujanić Marija</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-europeana_collections_9200447_BibliographicResource_30000955430023</frbrgroupid><rsrctype>dissertations</rsrctype><prefilter>dissertations</prefilter><language>srp</language><creationdate>2012</creationdate><topic>diagnostics</topic><topic>dijagnostika</topic><topic>genotipizacija</topic><topic>genotyping</topic><topic>glodari</topic><topic>human material</topic><topic>humani materijal</topic><topic>molecular detection</topic><topic>molekularna detekcija</topic><topic>parasite strains</topic><topic>parazita</topic><topic>PCR</topic><topic>PCR-RFLP</topic><topic>rodents</topic><topic>sojevi</topic><topic>toksoplazmoza</topic><topic>Toxoplasma gondii</topic><topic>toxoplasmosis</topic><toplevel>online_resources</toplevel><creatorcontrib>Vujanić Marija</creatorcontrib><collection>Europeana Collections</collection></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext_linktorsrc</fulltext></delivery><addata><au>Vujanić Marija</au><format>dissertation</format><genre>dissertation</genre><ristype>THES</ristype><Advisor>Ivović Vladimir</Advisor><Advisor>Savić-Pavićević Dušanka</Advisor><Advisor>Đurković-Đaković Olgica</Advisor><btitle>Molecular detection and genotyping of Toxooplasma gondiistrains isolated in Serbia</btitle><date>2012-07-09</date><risdate>2012</risdate><abstract>Toxoplasmosis, infection caused by the parasite Toxoplasma gondii, is one of
the most widespread zoonoses in the world and it has been estimated that up
to one third of the global human population is infected with this parasite.
Although serological tests are the primary method in the routine diagnosis of
toxoplasmosis, interpretation of the results can be not only difficult and
time-consuming, but they can be inconclusive. Therefore, the introduction of
highly sensitive molecular methods, by which results can be obtained within a
day, into the diagnosis of toxoplasmosis is of great importance. The aim of
this research was the introduction of molecular methods into the diagnosis of
toxoplasmosis for the first time in Serbia, as well as genotypization of the
isolated T. gondii strains. This study included 160 human biological samples
from 127 patients serologically suspected of toxoplasmosis in which the
presence of T. gondii was analyzed by PCR and by bioassay in mice. It was
found that the detection of T. gondii DNA in human biological materials
depends on the type of sample as well as on the initial volume. Real-time PCR
was positive in 30.8% of peripheral blood samples, 33.3% of cord blood
samples, 35.7% of amniotic fluid samples, 57.1% of CSF samples and 60% of
humor aqueous samples. The detection rate of T. gondii by molecular methods
was higher than by bioassay, in both blood samples from adults suspected of
acute toxoplasmosis and samples from fetuses and newborns suspected of
congenital toxoplasmosis. The results of this study emphasize the huge
practical importance and potential of molecular diagnostics and show the
benefit of introducing this method as part of the standard protocol for the
diagnosis of toxoplasmosis. T. gondii has a wide range of host organisms,
including all warm-blooded animals, so that in the second part of this
research molecular methods for the detection of this parasite were applied in
animal material, rodents specifically. Rodents were chosen because of their
potentially important role as a link in the transmission chain of T. gondii
infection. A total of 156 brain samples from 144 brown rats (Rattus
norvegicus) and 12 house mice (Mus musculus) were analyzed. T. gondii DNA was
vi detected in 10.4% of rats and in as many as 83.3% of house mice. The rate
of positive findings in rats and mice collected in Belgrade is an indicator
of the degree of contamination of the urban environment with T. gondii,
reflecting a considerable public health risk. Finally, this research provides
the very first and original phylogenetic data on T. gondii population
structure in Serbia. Recent studies performed world-wide indicate the
existence of a clonal population structure characterized by three main,
widespread clonal lineages (I, II and III), of which types I and II are
predominant in Europe and North America. There are also recombinant and
atypical strains that are mainly present in South America and Africa. In this
study, the identification of the T. gondii clonal type was performed in 22
samples, 19 from humans and three from rats, using the PCRRFLP method.
Genotyping was successful in five human samples of which four isolates
originating from cases of congenital toxoplasmosis, toxoplasmosis in
pregnancy and bone marrow transplantation were type II, while one isolate
from a case of congenital toxoplasmosis was type I. These results, first for
the Southeastern Europe, indicate the predominance of genotype II in our
country, as well as in most of the Europe.
Toksoplazmoza, oboljenje koje izaziva parazit Toxoplasma gondii, jedna je od
najrasprostranjenijih zoonoza u svetu, i procenjuje se da je čak jedna
trećina čovečanstva inficirana ovim parazitom. Iako serološki testovi
predstavljaju primarnu metodu u rutinskoj dijagnostici toksoplazmoze,
interpretacija rezultata seroloških analiza u nekim kliničkim slučajevima
može biti komplikovana i dugotrajna, a neretko i nedovoljno informativna.
Zbog toga je uvođenje molekularnih metoda koje inače odlikuje velika
osetljivost i pomoću kojih je moguće dobiti rezultate u kratkom vremenskom
periodu, od velikog značaja za dijagnostiku toksoplazmoze. Otuda je cilj ovog
istraživanja bio uvođenje molekularne dijagnostike toksoplazmoze po prvi put
u Srbiji, kao i genotipizacija izolovanih sojeva T. gondii. Ovom studijom je
obuhvaćeno 160 uzoraka poreklom od 127 pacijenata serološki suspektnih na
toksoplazmozu, kod kojih je prisustvo T. gondii analizirano PCR-om u realnom
vremenu i biološkim ogledom. Pokazano je da uspešnost dokazivanja DNK T.
gondii u materijalu humanog porekla zavisi kako od tipa materijala tako i od
ukupne zapremine materijala od kojeg se uzima uzorak. Tako je PCR u realnom
vremenu bio pozitivan u 30,8% uzoraka periferne krvi, u 33,3% uzoraka krvi iz
pupčanika, u 35,7% uzoraka plodovih voda, u 57,1% uzoraka likvora i u 60%
uzoraka očnih vodica. U odnosu na izolaciju parazita biološkim ogledom,
pokazana je znatno viša stopa detekcije DNK T. gondii, i to i u uzorcima krvi
od pacijenata suspektnih na akutnu toksoplazmozu tako i u onima poreklom od
fetusa i novorođenčadi suspektnih na kongenitalnu toksoplazmozu. Dobijeni
rezultati ukazuju pre svega na veliki potencijal i praktični značaj
molekularne dijagnostike kao i na potrebu uvođenja ove metode kao dela
standardnog protokola za dijagnostiku toksoplazmoze. T. gondii ima veoma
širok spektar domaćina koji uključuje sve toplokrvne životinje, te su u drugom
delu ovog istraživanja molekularne metode za detekciju toksoplazmoze primenjene
na materijalu životinjskog porekla, i to konkretno od glodara. Zbog njihove
potencijalno značajne uloge kao karike u epidemiološkom lancu toksoplazmoze u
okviru ove studije ispitano je i 156 uzoraka poreklom od 144 braon pacova
(Rattus norvegicus) i 12 kućnih miševa (Mus musculus). DNK T. gondii je
detektovana u mozgu 10,4% pacova i kod čak 83,3% kućnih miševa. Relativno
visoka stopa pozitivnih nalaza kod pacova i miševa sakupljenih u Beogradu
ukazuje na kontaminiranost naše sredine parazitom T. gondii koja predstavlja
rizik za javno zdravlje. Poslednji deo ovog rada se odnosi na prva i poptuno
originalna filogenetska istraživanja populacione strukture T. gondii. Naime,
skorašnja istraživanja u svetu ukazuju na postojanje klonske populacione
strukture koju karakterišu tri široko rasprostranjena glavna klonska tipa (I,
II i III), od kojih su tipovi I i II predominantni u Evropi i Severnoj
Americi. Postoje takođe i rekombinantni i atipični sojevi koji su uglavnom
prisutni u Južnoj Americi i Africi. U ovom istraživanju određivanje klonskog
tipa, čime su dobijeni i prvi podaci o populacionoj strukturi T. gondii u
Srbiji, vršeno je korišćenjem PCR-RFLP metode. Analizirano je ukupno 22 uzorka
i to 19 humanog porekla i 3 poreklom od pacova. Uspešno je genotipizirano pet
uzoraka poreklom od ljudi, za koje se pokazalo da četiri pripadaju tipu II a
jedan tipu I. Sojevi tipa II su izolovani iz kliničkih uzoraka bolesnika sa
kongenitalnom toksoplazmozom, toksoplazmozom stečenom u trudnoći i
reaktivacijom infekcije usled post- transplantacione imunosupresije, dok je
soj tipa I izolovan iz jednog uzorka poreklom od kongenitalno inficiranog
novorođenčeta. Ovi rezultati, prvi za čitavo područje Jugoistočne Evrope,
pokazuju da je i u našoj zemlji, kao i u većem delu Evrope, predominantan pre
svega genotip II.</abstract><pub>University of Belgrade, Faculty of Biology</pub><oa>free_for_read</oa></addata></record> |
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| recordid | cdi_europeana_collections_9200447_BibliographicResource_3000095543002 |
| source | Europeana Collections |
| subjects | diagnostics dijagnostika genotipizacija genotyping glodari human material humani materijal molecular detection molekularna detekcija parasite strains parazita PCR PCR-RFLP rodents sojevi toksoplazmoza Toxoplasma gondii toxoplasmosis |
| title | Molecular detection and genotyping of Toxooplasma gondiistrains isolated in Serbia |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-04T13%3A38%3A47IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-europeana_1GC&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft.genre=dissertation&rft.btitle=Molecular%20detection%20and%20genotyping%20of%20Toxooplasma%20gondiistrains%20isolated%20in%20Serbia&rft.au=Vujani%C4%87%20Marija&rft.date=2012-07-09&rft_id=info:doi/&rft_dat=%3Ceuropeana_1GC%3E9200447_BibliographicResource_3000095543002%3C/europeana_1GC%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_id=info:pmid/&rfr_iscdi=true |