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Detection of Brugia malayi microfilaria/Larvae in mosquito using Polimerase Chain Reaction. Haryuningtyas, Dyah; Subekti, Didik Tulus
Indonesian Journal of Animal and Veterinary Sciences Vol 13, No 3 (2008)
Publisher : Indonesian Animal Sciences Society

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (203.014 KB) | DOI: 10.14334/jitv.v13i3.587


Lymphathic filariasis that is also known as elepanthiasis is caused by infestation of 3 species nematode Wuchereria bancrofti, Brugia malayi and Brugia timori. In Indonesia 70% filariasis case caused by Brugia malayi. Mosquito species from genus Anopheles, Aedes, Culex, Mansonia and Armigeres are known as vector of this disease. Microfilaria detection on mosquito is one methode to know infection rate in vector population in endemic area.The objectives of the research were to study the ability of Hha1 repeat applicable to detect microfilaria/larvae in a pool of mosquitoes and to get description of adult mosquito night biting population lived in endemic area of filariasis brugian. Mosquito as positive control used in this research come from laboratory of parasitology of FKUI. Mosquito sample from the field was from Binawara and Kolam Kiri villages, South Kalimantan province. Mosquito were trapped then identified by its species. DNA of mosquitoes was extracted and then run by the PCR using Hha 1 repeat primer. Result of the research indicated that adult mosquitoes night biting from Binawara village consist of Culex, Mansonia, Anopheles genus and from Kolam Kiri village only from Mansonia genus. Hha 1 repeat primer is applicable to detect 1 mosquito infected with microfilaria/larvae in a pool of negative mosquitoes. Mosquito samplesfrom the two villages showing negative PCR.   Key Words: Filariasis, Brugia Malayi, Vector, Microfilaria, Filaria Larve, PCR
Cloning Gene Encoding Micronema 3 (Mic3) Protein of Tachyzoite Toxoplasma Gondii Local Isolate Artama1, Wayan T.; Dewi, Ni Nyoman Ayu; Subekti, Didik Tulus
Indonesian Journal of Biotechnology Vol 10, No 1 (2005)
Publisher : Universitas Gadjah Mada

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Microneme 3 (MIC3) protein tachyzoites Toxoplasma gondii is one of protein which plays an important roleduring cell host invasion. Gene encoding MIC3 protein has been studied and it was suggested a potent vaccinecandidate against Toxoplasma gondii infection. The aim of this research is to clone and sequence the gene encodingMIC3 protein of tachyzoites Toxoplasma gondii local isolate by amplification using polymerase chain reaction withspecific primers. The amplified DNA fragment was cloned into pGEM-T and transformed into E. coli XL-1 Blue byheat shock method. Recombinant plasmids were isolated using alkali lysis method and analyzed by digestionusing restriction endonuclease enzymes PstI, HindIII, NcoI and EcoRV. The recombinant plasmids then sequencedto find out the nucleotide sequence of insert gene by ABIPRISM 377 DNA Sequencer. The DNA sequence thenwere analyzed by computer software for alignment. The result showed that transformation in E. coli XL-1 Blue bypGEM-T produced one clone that was encoding MIC3 protein. Analysis of 489 bp from 5’ and 447 from 3’ of genesequence showed 97-98% homology with gene encoding for MIC3 protein of RH isolate.Keywords: MIC3 protein, Toxoplasma gondii, tachyzoite, recombinant DNA
Cloning and Sequencing cDNA Encoding for Rhoptry-2 Toxoplasma Gondii Tachyzoite Local Isolate Artama, Wayan T.; Sari, Yulia; Subekti, Didik Tulus; Poerwanto, Soenarwan Hery; Subandono, Jarot
Indonesian Journal of Biotechnology Vol 10, No 2 (2005)
Publisher : Universitas Gadjah Mada

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Rhoptry protein belongs to an excretory and secretory antigens (ESAs) that play an important role during activepenetration of parasite into the cell target. This protein an able Toxoplasma gondii to actively penetrate targetedcell, meanwhile ESAs protein stimulates intracellular vacuole modification. It is, therefore, after the parasitesuccessfully enter the cell target then Granule (GRA) proteins are responsible for the formation of parasitophorusvacuole, which is protect the fusion with other intracellular compartments such as lysosomal vacuole. Consequently,this parasite is being able to survive and multiply at the cell target. The current study was aimed to clone andsequens cDNA encoding for ROP-2 of local isolated T. gondii tachizoite through DNA recombinant technique.Total ribonucleic acid (RNA) was isolated from tachyzoites of local isolated T. gondii that were grown up in Balb/c mice. Messenger RNA was isolated from total RNA using PolyAtract mRNA Isolation System. Messenger RNA wasused as a template for synthesis cDNA using Riboclone cDNA Synthesis System AMV-RT. EcoRI adaptor fromRiboclone EcoRI Adaptor Ligation System was added to Complementary DNA and than ligated to pUC19. Recombinantplasmid was transformed into E. coli (XL1-Blue). The transformed E. coli XL-1 Blue were plated on LB agarcontaining X-Gal, IPTG and ampicillin. Recombinant clones (white colony) were picked up and grown up in theLB medium at 37oC overnight. Expression of recombinant protein was analysed by immunoblotting in order toidentify cDNA recombinant wich is express ESA of T. gondii local isolate. Recombinant plasmid were isolatedusing alkalilysis method and were elektroforated in 1% agarose gel. The isolated DNA recombinant plasmid wascut using Eco RI and then sequenced through Big Dye Terminator Mix AB1 377A Sequencer using M13 Forward andM13 Reverse primers. The conclusion of this results showed that the recombinant clone was coding for excretoryand secretory protein which has molecular weight of 54 kDa. The DNA alignments of sequence from the clonedgene showed 97% homology with gene encoding for ROP-2 of T. gondii RH isolate.Keywords: Toxoplasma gondii, tachizoite, ESA, complementary DNA, ROP2
BERITA BIOLOGI Vol 4, No 4 (1998)
Publisher : Research Center for Biology-Indonesian Institute of Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14203/beritabiologi.v4i4.1265


Study on the comparison between allantoin (5 ¢ ureidohydantoin) and Betadine ® (povidone iodine) was conducted to compare and evaluate their efficacy, especially to accelerate wound (incision) healing. Treatment divided into three groups, first group is Control (without therapy), second group is allantoin treatment and the last one is Betadine ® treatment. Allantoin obtained from cattle's urine by Meissner method. The solution made of 2,4 grams of allantoin in 600 milliliters aqueous solution. Treatments (therapies) were given three times a day topically. Results showed no significant difference between allantoin and Betadine ® treatments (p > 0,05), control and the other treatments i.e allantoin and Betadine ® therapies have significantly difference (p < 0,01).