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ANALISIS MOLEKULER PROFIL PROTEIN PILLI UNTUK MENGUNGKAP HUBUNGAN SIMILARITAS 26 STRAIN SALMONELLA TYPHI ISOLAT JAWA Darmawati, Sri; Haribi, Ratih; Anwar, Syaiful
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2012: SEMINAR NASIONAL HASIL PENELITIAN 2012
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Variasi dan hubungan similaritas 26 strain Salmonella typhi Isolat Jawa merupakanawal untuk melacak protein sub unit pilli spesifik yang memiliki aktivitas hemaglutinasi.Tujuan penelitian analisis molekuler profil protein pilli untuk mengungkap hubungansimilaritas 26 Strain S. typhi Isolat Jawa.Analisis dilakukan terhadap 26 strain yang berasal dari Surabaya, Madiun,Malang, Salatiga, Magelang, Bandung, Bogor, Jakarta, Yogyakarta dengan elektroforesisSDS-PAGE. Analisis hubungan similaritas digunakan program MVSP, untukmengkonstruksi dendogram yang mencerminkan klasifikasi dari 26 strain S. typhi IsolatJawa berdasarkan nilai indeks similaritas (S SM ) dengan algoritma UPGMA.Hasil analisis profil protein pili menunjukkan (1) Jumlah pita protein sub unitpilli bervariasi : 8-17 pita, BM tertinggi 200 kD, terendah 10 kD, dengan 20 karakter. (2)Protein 100 kD, 50 kD, 45 kD dan 40 kD adalah protein sub unit pilli yang dimiliki oleh26 strain S. typhi Isolat Jawa. (3) Dari 26 strain S. typhi Isolat Jawa terdiri dari duakelompok besar yang mempunyai indeks similaritas 61,2%. Kelompok pertama adalahstrain S. typhi Isolat Jawa Tengah dan Jawa Timur, dan kelompok ke dua adalah strain S.typhi Isolat Jawa Barat dan DKI. Pita 14 (12,5 kD) dan 15 (78kD) dari protein sub unitpilli hanya dimiliki strain S. typhi dari Surabaya. Pita 18(35kD) dan 20 (72kD) dariprotein sub unit pilli hanya dimiliki oleh Strain S. typhi dari DKI Jakarta.
DETEKSI GEN COA PADA STAPHYLOCOCCUS AUREUS YANG DIISOLASI DARI SUSU SAPI MURNI Wahyuni, Rizka Ayu; Darmawati, Sri; Prastiyanto, Muhammad Evy
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2017: Prosiding Seminar Nasional Publikasi Hasil-Hasil Penelitian dan Pengabdian Masyarakat
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Tujuan penelitian ini untuk mendeteksi gen Coa pada S. aureus yang diisolasi dari susu sapi murni di Kelurahan Gedawang. Jenis penelitian ini adalah penelitian deskriptif. Bakteri S. aureus di isolasi dari 15 sampel susu sapi murni. Deteksi gen Coa S. aureus menggunakan primer spesifik  Forward (5´-ATAGAGATGCTGGTACAGG-´3) dan primer Coa Reverse (5´GCTTCCGATTGTTCGATGC-3´).Jumlah isolat S. aureus yang didapatkan dari Isolasi Bakteri sebanyak 3 sampel positif yang memiliki gen Coa dengan hasil produk 756 bp. Hasil pada penelitian ini menunjukkan hasil positif gen Coa sebesar 100%.   Keywords: Staphylococcus aureus, Susu, gen Coa
ANALISIS PROTEIN PILLI SALMONELLA TYPHI LSOLAT RS. KARIADI SEMARANG DENGAN ELEKTROFORESIS SDS-PAGE Darmawati, Sri; Haribi, Ratih
JURNAL LITBANG Vol 2, No 3 (2005): Penelitian, Pengembangan, dan Pengabdian
Publisher : JURNAL LITBANG

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Abstract

Background : Salmonella typhi is bacteria which causes typhoid fever. Typhoid fever is severally serious, infectious decease and is endemic decease in Indonesia with relatively high frequency rate around 358-810/100.000 people each year This decease is usually spreading along with many kinds of decease which also have relatively high mortality rate, that is 1-5% of the sufferers (Punjabi, NH. , 2 004) . The infection process of bacteria into human body is signified with bacteria Cell adhering in mucosa intestinal surface. Pilli is structured by pilli protein consisting of Several sub units of pilli protein Objective: conducting pilli protein Salmonella typhi isolate Kariadi Hospital Semarang Analysis through electrophoreses SDS-PAGE, so that take amount of sub unit of pilli protein with each its molecule weight can be found out. Method : This research is conducted through three stages; firstly, bacteria cultivation with Biphasic media (BHI Agar and broth BHI ); secondly, pilli isolation with Ehara method (/956); Thirdly, sub unit of pilli protein separation with I 2% SDS-PAGE based on Lemmli method (1970). Result: There are four major sub units of protein in pilli, that is, 36kDa; 26,5 kDa; 22,2kDa; 18,6lrDq and there are also four minor sub units of protein pilli, that is, 116 kDa; 62,3kDa;45kDa;20,9kDa,andseveral other minor sub units of protein with every thin band. Keywords : Salmonella and Electrophoresis
MOLECULAR CHARACTERIZATION AND HEMAGGLUTINATION ACTIVITIES OF FLAGELLIN PROTEIN OF SALMONELLA TYPHI Darmawati, Sri; Santosa, Budi; Prastiyanto, Muhammad Evy; Saptaningtyas, Ragil
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2016: Proceeding of International Seminar on Education Technology (ISET) 2016
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Abstract. The purposes of this research are for molecular characterization and hemagglutination activity test of flagellinprotein of Salmonella typhi. The research samples consist of 7 strains of S. typhi isolates from Central Java (5 strains from Semarang city, 1 strain from Salatiga and 1 strain from Magelang) and 2 strains of S. typhi from Yogyakarta (Doctor Sardjito Hospital and Bethesda Hospital). The undertaking procedures are: 1) PCR and sequencing of fliC genes using primer LPW 1856 and LPW 1857.2) Isolation and separation of flagellin protein using SDS-PAGE. 3) Hemagglutination Activity Test upon human erythrocytes of blood group A, B, AB and O.The results show that 8 strains of S. typhi have a fliC gene size of 1452 to 1488 bp including serovar H1-d, and 1 strain with the size of 1267 bp including serovar H1-J. Flagella protein resulted from SDS-PAGE protein consists of 1-2 major proteins and 1-3 minor proteins with a molecular weight of 16-116 kDa. The results of hemagglutination activity test of flagellin protein show that there are 3 strains of S. typhi (MG-1, SA02.2 and BET) which are able to agglutinate human erythrocytes of bloodgroup A, B, AB and O (2-64HA), 6 other strains show various hemagglutination activities varied
HAEMAGGLUTINATION ACTIVITY OF SALMONELLA TYPHI FLAGELLIN PROTEIN BASED ON ABO BLOOD GROUP Saptaningtyas, Ragil; Darmawati, Sri; Dewi, Sri Sinto
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2015: Prosiding Student Paper Presentation The 2nd University Research Colloquium
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Salmonella typhi is a negative basil Gram bacterium that causes typhoid fever. Flagel of S. typhi contain proteins, as locomotor and it can help bacteria to attach on host cells. Haemagglutinin protein is protein that can agglutinate erythrocytes. The goal of this research is to analize S. typhi flagellin proteins haemagglutination activity Based on ABO blood group. Flagellin proteins isolation method of modified Alexan method (2009) and haemagglutination test method of Finkeltein and Hanne method (1982). Haemagglutination result of ABO blood type show that SLT-1 S. typhi flagellin proteins can?t agglutinate human erythrocytes A, B, and O, but it can agglutinate human erythrocytes until 8 times dilution of AB blood type from 50 ?g/?l concentration in 50 ?l. BA 07.4 S. typhi flagellin proteins can agglutinate human erythrocytes of A blood type until 16 times dilution, 8 times dilution of AB blood type, 4 times dilution of O blood type, and it can?t agglutinate human erythrocytes B blood type.Keywords: Salmonella typhi, Flagellin Proteins, Haemagglutination, Erythrocytes of ABO System.
ISOLASI DAN IDENTIFIKASI MOLEKULER BAKTERI PENGHASIL ENZIM PROTEASE PSEUDOMONAS STUTZERI ISTD4 DARI TEMPE GEMBUS PASCA FERMENTASI 1 HARI Inayatul, Wa Ode; Muchlissin, Sakti Imam; Mukaromah, Ana Hidayati; Darmawati, Sri; Ethica, Stalis Norma
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Proteaseis  a group  of enzymes that  play  an important  role in  biochemical reactions, whichc a use protein break down. Protease is among main enzymes used in industry, which commercial value reach 60% of total enzymes world wide. This study a imed toisolat protease-producing bacterium found on tempe gembus in after 1-day post-fermentation and to identify the bacterial isolat obtained based on the analysis of its 16S rRNA gene. Isolati on and purification process wasd one using Nutrient Agar media with spread technique. The protease production test was carried out on skim milk agar medium. The molecular identification process was performed by analyzing sequence of 16S rRNA gene fragment of bacteria amplified using both forward primer F (F:5'- AGAGTTGATCCTGGCTCAG-3'), and reverse primer R (R:5'- GGTTACCTTGTTACGACTT-3') by Polymerase Chain Reaction (PCR) method. The amplified DNA from PCR was then sequenced. From the isolation process a bacterial strain that has a proteolytic activity based on observation of clear zone area with a diameter of 85 mm was obtained. From sequence alignment result using BLAST (Basic Local Alignment Search Tool) the fragment of 16S rRNA gene of strain ISTD1.4 obtained has similarity level of 98% with fragment of 16S ribosomal RNA gene of bacterium Pseudomonas stutzeri strain E141. In conclusion, strain ISTD1.4 is a potential protease-producing bacteria and is identified as Pseudomonas stutzeri ISTD4. Keywords: Bacterial isolation, molecular identification, proteolytic bacteria, 16S rRNA gene
DETECTION OF VIRULENCE GENES PHOP AND PHOQ IN SALMONELLA SPP. USING IN SILICO POLYMERASE CHAIN REACTION Ethica, Stalis Norma; Fuad, Hayatun; Hidayah, Nur; Dewi, Sri Sinto; Ernanto, Aditya Rahman; Sulistyaningtyas, Ayu Rahmawati; Silvestre, Richard David; Darmawati, Sri
RESEARCH FAIR UNISRI Vol 4, No 1 (2020): RESEARCH FAIR UNISRI
Publisher : RESEARCH FAIR UNISRI

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Abstract

Detection of Salmonella bacteria based on their virulence genes is among essential steps in the eradication of clinical infection by bacteria. In this study, two pair of primers, PhoPF-PhoPR: 5?- CCGCGCAGGAAAAACTCAAA-3? and 5?-ATCTGTTCCAGCATCACCGG -3? as well as PhoQF-PhoQR: 5?-AGAGATGATGCGCGTACTGG-3? and 5?- CAGACGCCCCATGAGAACAT-3?, had been successfully designed using Primer3Plus to detect the presence of phoP and phoQ genes in Salmonella spp. Using genomic DNA of 44 genomic data of Salmonella spp. as templates, PhoPF-PhoPR could produce 520-bp amplicon, while PhoQF-PhoQR could result in 598-bp amplicon. Results of in silico PCR showed that both pairs of primers PhoPF-PhoPR and PhoQF-PhoQR could detect only Salmonella enterica species, and no Salmonella bongori species could be detected based on phoP and phoQ sequences. Both pairs of PhoPF-PhoPR and PhoQF-PhoQR primers were also able to detect the virulence genes in most of the studied subspecies of Salmonella enterica available in silico database unless Arizona subspecies. As conclusion, based on this in silico study, phoP and phoQ genes appeared to be biomarkers for Salmonella enterica species. Both pairs of primers designed in this study has potential to be used as detection tool to differentiate species Salmonella enterica from Salmonella bongori, and also to distinguish S.enterica subsp. enterica from subsp. Arizonae.Keywords: Gene detection, bacterial virulence, phoP, phoQ, Salmonella spp.
ISOLASI BAKTERI PENGHASIL ENZIM PROTEASE BACILLUS AMYLOLIQUEFACIENS IROD2 PADA ONCOM MERAH PASCA FERMENTASI 48 JAM Pamaya, Dwi; Muchlissin, Sakti Imam; Maharani, Endang Tri Wahyuni; Darmawati, Sri; Ethica, Stalis Norma
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Proteolytic  bacteria  are  the  bacteria  capable  of  producing  extracellular protease enzymes, namely protein-breaking enzymes that are widely used in many industrial fields. This study aimed to isolate a proteolytic bacterium found on 48-h post-fermented oncom and molecular identification method. The initial isolation and purification process of the colony was carried out using Nutrient Agar medium. Selection of protease enzyme obtained by bacterial isolate was done on Milk Skim Agar medium. Identification process of the isolate was done through amplification of 16S rRNA gene using PCR, sequencing and analysis of gene sequences using BLAST program. From the isolation process  a bacterial isolate that has proteolytic by the ability to produce a clear zone of 82.00 on plate. The result of the 16S rRNA gene sequence analysis showed that the proteolytic bacterial isolate obtained in this study had a 98% homology level with 16S ribosomal RNA isolate of Bacillus amyloliquefaciens strain A1142 (Genbank access code: KTT722836.1). Based on the results of the molecular identification, the isolate was identified as Bacillus amyloliquefaciens strain IROD2  (IROD2 = Indonesia Red Oncom Day2). As conclusion, from 48-h post fermented red oncom, a protease producing bacterial strain molecularly identified as Bacillus amyloliquefaciens strain IROD2. Keywords: Moleculary was identified, Proteolitic bacteria, 16S rRNA gene
AKTIVITAS ANTIBAKTERI MADU TERHADAP BAKTERI MULTI DRUG RESISTANT SALMONELLA TYPHI DAN METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS Panjaitan, Rahma Asriani; Darmawati, Sri; Prastiyanto, Muhammad Evy
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2018: SEMINAR NASIONAL PENDIDIKAN SAINS DAN TEKNOLOGI
Publisher : Universitas Muhammadiyah Semarang

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Abstract

This study aimed to analyze the antibacterial activity of Palm oil Honey against Multi  Drug  Resistant  Salmonella  typhi  and  Methicillin-Resistant Staphylococcus aureus by concentration 50%, 60%, 70%, 80%, 90% and 100%. This  research  was  an  experimental  test  with  Posttest  Only  Control  Group Design in an in vitro manner using well diffusion method techniques. The well diffusion method used MHA which made by the well of 5 mm diameter and inserted 200 µ L sample then incubated 35 ± 20C for 24 hours. The results of the study showed the antibacterial activity of Palm oil Honey with zone inhibition against MDR S.typhi 11.4 mm (concentration 90%), 13.4 mm (concentration 100%) and against MRSA 11.7 mm (concentration 100%). The results of zone inhibition of Palm oil honey with concentration 100% against MDR S.typhi was larger than the  zone inhibition  against MRSA. The palm oil honey inhibition zone  against  S.typhi  MDR  bacteria  compared  with  sulfamethoxazole  (SXT) antibiotics with 25 mm inhibition zones was included in the intermediate category and the palm oil honey inhibition zone against MRSA bacteria compared with Tetracycline (TE) antibiotics with 23 mm inhibition zone was included in resistant category. Keywords: Antibacterial Activity, MDR S. typhi, MRSA, Honey, Inhibition Zone
DAYA HAMBAT EKTRAK ETANOL BUAH BELIMBING WULUH (AVERRHOA BILIMBI L) TERHADAP PERTUMBUHAN STAPHYLOCOCCUS AUREUS DAN STAPHYLOCOCCUS EPIDERMIDIS SECARA IN VITRO Rahmiati, Asri; Darmawati, Sri; Mukaromah, Ana Hidayati
PROSIDING SEMINAR NASIONAL & INTERNASIONAL 2017: Prosiding Seminar Nasional Publikasi Hasil-Hasil Penelitian dan Pengabdian Masyarakat
Publisher : Universitas Muhammadiyah Semarang

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Abstract

Acne infection is caused inflammation of pilosebasea accompanied by accumulation of keratin material, caused by S. aureus and S. epidermidis bacteria. The community uses of wuluhstarfruit (Averrhoabilimbi L) as a traditional medicine to treat acne infection. Wuluh starfruit contains flavonoids, alkaloids, tannins, and saponins that act as anti microbial. The aim of this study was to analyze the inhibitory power of wuluh starfruit ethanol extract on the growth of S. aureus and S. epidermidis bacteria. The method used in this research is the diffusion of wells. This research used two types of bacteria. S.aureus and S.epidrmidis, each bacteria of the four treatment groups that is 10%w/v; 20%w/v; 30%w/v; 40%w/v; positive control of Ciprofloxacin, and negative control of sterile aquades. The research results of inhibitory power of ethanol extract of wuluh starfruit with variation of concentration 10 %w/v; 20 %w/v; 30 %w/v; and 40 %w/v successively in S.aureus was 21.6 mm; 27.0 mm; 31.3 mm; And 34.0 mm, whereas in S.epidermidis is 28.6 mm; 31.6 mm; 36.3 mm; And 39.0 mm. Then the positive control of Ciprofloxacin has an inhibit zone of 30.0 mm and 35.0 mm. While the negative control of sterile aquades is not formed inhibit zone. The result ofOne Way Anova statistic test on S. aureus is p=0.000 and S. epidermidis is p=0.000, because (p<0.05) hence result there is significant difference, so it can be concluded that the extract of wuluh starfruit ethanol can inhibit the growth of S. aureus and S. epidermidis and there is a significant difference between the variantconcentration ofethanol extract wuluhstarfruit. Keywords: Staphylococcus aureus, Staphylococcus epidermidis, Ethanol extract of wuluhstarfruit.