scholarly journals Eliminasi Endosimbion Wolbachia sp. pada Nyamuk Aedes albopictus dengan Antibiotik Tetrasiklin

2021 ◽  
pp. 171-178
Author(s):  
Endang Srimurni Kusmintarsih ◽  
Darsono Darsono ◽  
Edy Riwidiharso ◽  
Rokhmani Rokhmani ◽  
Trisnowati Ambarningrum ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Hemorrhagic Fever ◽  
Survey Method ◽  
Chain Reaction ◽  
Specific Primers ◽  
Tetracycline Antibiotics ◽  
Sugar Feeding ◽  
Pcr Method ◽  
Primary Vector ◽  
Polymerase Chain

Dengue Hemorrhagic Fever (DHF) is a disease caused by the dengue virus which is transmitted through the bite of the Aedes mosquito. Aedes aegypti, which is known as the primary vector of dengue virus, is naturally not infected by Wolbachia sp. endosymbiont, while Ae. albopictus which is a secondary vector naturally infected with Wolbachia sp. The Wolbachia sp. known to inhibit the transmission of Dengue virus, to study the mechanism, it is necessary to eliminate Wolbachia sp. from Ae. albopictus, then infects the Ae. albopictus with the Dengue virus. The aim of the study was to determine the ability of tetracycline antibiotics to eliminate Wolbachia sp. from the Ae. albopictus mosquito. Ae. albopictus eggs was obtained in the Ciamis area by survey method using ovitrap which was installed outside the house. The Ae. albopictus eggs are then incubated in the laboratory and reared until they become adult mosquitoes. Mosquitoes were treated with sugar feeding which had been given tetracycline 0.25mg/ml every two days alternated with blood feed. Detection of the presence of Wolbachia sp. on mosquitoes carried out in first to third generations by the polymerase chain reaction (PCR) method using Wsp-specific primers. The results showed that the first to third generation mosquitoes were still infected with Wolbachia sp. This shows that the dose of tetracycline antibiotics used has not been able to eliminate Wolbachia sp. from the Ae. albopictus mosquito.

scholarly journals Precipitation Solvents for RNA Extraction of Dengue Virus Type 3: Dimethylformamide, Ethylenediamintetraacetic Acid, and Ultrapure H2O

2019 ◽  
Vol 3 (2) ◽  
pp. 78
Author(s):  
Rizqidhana Juliana Putri ◽  
Teguh Hari Sucipto ◽  
Harsasi Setyawati ◽  
Siti Churrotin ◽  
Ilham Harlan Amarullah ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Dengue Fever ◽  
Reverse Transcription ◽  
Rna Extraction ◽  
Chain Reaction ◽  
Dengue Virus Type 3 ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Type 3

Dengue is a disease caused by a virus from the family Flaviviradae, carried by a female mosquito of Aedes aegypti species. Dengue fever is widespread in the tropic areas. It caused by rainfall, temperature and unplanned urbanization. According to the ministry of health , almost all provinces in Indonesia are endemic areas of dengue fever. In 2014, up to mid-December Dengue Hemorrhagic Fever (DHF) patients in 34 provinces in Indonesia are 71,668 people and 641. This figure is lower than the previous year, 2013 with 112,511 people and 871 deaths . This disease consists of four types of serotypes, namely DENV-1, DENV-2, DENV-3, and DENV-4. This disease can be identified using a variety of methods, one of the method is Reverse Transcription - Polymerase Chain Reaction (RT-PCR) method. This study aims to determine the ability of Dimethylformamide (DMF), Ethylenediamintetraacetic Acid (EDTA), and Ultrapure H2O as the substitute of  Ethanol for precipitation in RNA extraction process. The sample used in this research obtained from Surabaya. RNA extraction itself can be done by using a special kit for RNA extraction. In Reverse Transcription - Polymerase Chain Reaction method, first RNA is extracted and then transcribed back (Reverse Transcription) which then form cDNA that later will be amplified by using PCR method. In this study used specific primers for dengue virus type 3 (DENV-3). The results of this study show that DMF, EDTA, and Ultrapure H2O can be used as the substitute of Ethanol for precipitation on RNA extraction. The result is evidenced by the formation of viral DNA bands on gel electrophoresis results.

scholarly journals Mite Sarcoptes scabiei Varieties Hominis in South Sumatra: Specific Identification and Comparative Study

2020 ◽  
Vol 8 (A) ◽  
pp. 938-942
Author(s):  
Yessi Arisandi ◽  
Chairil Anwar ◽  
Salni Salni ◽  
Dadang Hikmah Purnama ◽  
Novrikasari Novrikasari ◽  
...  
Keyword(s):  
Deoxyribonucleic Acid ◽  
Sarcoptes Scabiei ◽  
Chain Reaction ◽  
Specific Primers ◽  
Specific Identification ◽  
16S Gene ◽  
Dna Strands ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Relationship Of

BACKGROUND: Sarcoptes scabiei mites have more than 15 genetically diverse varieties from various hosts. Identification of S. scabiei mite varieties hominis as an intervention in its prevention is still rarely done. AIM: This study aimed to observe the genetic relationship of the mite S. scabiei varieties hominis compare to the parasite S. scabiei varieties hominis in other regions. METHODS: This research used polymerase chain reaction (PCR) and sequencing methods with 16S gene-specific primers. From 32 S. scabiei samples, 22 samples were identified as varieties hominis that was marked by the appearance of the band at 132 bp. RESULTS: S. scabiei mites hominis varieties from South Sumatra (Yessi Scabies A2 and Yessi Scabies B3) have similarities with deoxyribonucleic acid (DNA) strands with S. scabiei hominis varieties from China (KJ781377 and KJ781376). In contrast, Yessi Scabies A1 has similarities with DNA strands with mite S. scabiei varieties hominis from Australia (AY493402). Still, all the DNA strands, this research is different from S. scabiei mites DNA strands hominis from Panama and Pakistan. CONCLUSION: The PCR method is advantageous and specific in identifying mites S. scabiei hominis varieties, the cause of scabies in humans.

scholarly journals Isolation and Toxin Typing of Clostridium Perfringens From Sheep, Goats, and Cattle in Fars Province, Iran

2020 ◽  
Vol 8 (3) ◽  
pp. 89-93
Author(s):  
Masoumeh Hayati ◽  
Mehrdad Shamseddini ◽  
Yahya Tahamtan ◽  
Safar Sadeghzadeh ◽  
Mohsen Manavian ◽  
...  
Keyword(s):  
Clostridium Perfringens ◽  
Fars Province ◽  
Type B ◽  
Chain Reaction ◽  
Specific Primers ◽  
Defined Media ◽  
Different Types ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Published Research

Background: Clostridium perfringens is an important anaerobic bacterium found in the intestine of some livestock. It is concerned with the etiology of some diseases including enterotoxaemia. Various diseases are caused by different types of C. perfringens. Nonetheless, there is no published research on molecular typing and distribution of this pathogenic microorganism in Fars province. Objectives: Accordingly, our study focused on the isolation and toxin typing of C. perfringens from sheep, cattle, and goats in different parts of Fars province by the culture and the polymerase chain reaction (PCR) method. Materials and Methods: Approximately 459 fecal samples were collected and cultured on defined media for the isolation of C. perfringens. The confirmed isolates were genotyped by the PCR method using specific primers. Results: C. perfringens was isolated from 30.93% of the total samples. The results of toxin typing showed a total of 76 (54%), 13 (9%), 30 (21%), and 23 (16%) isolates as types A, B, C, and D, respectively. Conclusion: Our results indicated that C. perfringens type A was the most common type in sheep, cattle, and goats while the lowest number of isolates belonged to type B. Finally, the isolation of C. perfringens and toxin typing increase our knowledge of the epidemiology of these diseases and can help in the vaccine industry and better controlling related diseases.

A Multiplex PCR Method for the Identification of Listeria spp. and Listeria monocytogenes in Dairy Samples

1995 ◽  
Vol 58 (8) ◽  
pp. 867-872 ◽  
Author(s):  
LIEVE M. F. HERMAN ◽  
HERMAN F. M. DE RIDDER ◽  
GEERTRUI M. M. VLAEMYNCK
Keyword(s):  
Listeria Monocytogenes ◽  
Multiplex Pcr ◽  
Gene Sequence ◽  
Chain Reaction ◽  
Specific Primers ◽  
16S Rrna Sequence ◽  
Bacteriological Method ◽  
Agarose Electrophoresis ◽  
Pcr Method ◽  
Polymerase Chain

A multiplex polymerase chain reaction (PCR) method for the identification of Listeria spp. and Listeria monocytogenes has been developed. For the identification of Listeria spp. two specific primers were derived from the 16S rRNA sequence. For the identification of Listeria monocytogenes specific primers were derived from the gene sequence of the invasion-associated protein. The two amplified DNA fragments, respectively 1003 bp and 593 bp, were separated by agarose electrophoresis. The results obtained by the multiplex PCR method were in agreement with those obtained by the classical bacteriological method for the 64 dairy samples with presumptive positives.

scholarly journals PROFIL VIRUS DENGUE DI SURABAYA TAHUN 2008–2009

2018 ◽  
Vol 17 (1) ◽  
pp. 21
Author(s):  
Aryati . ◽  
Puspa Wardhani
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Hemorrhagic Fever ◽  
Human Pathogens ◽  
Virus Infections ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Who Criteria ◽  
Private Laboratory ◽  
Polymerase Chain

Four serotypes of dengue viruses (DENV) 1–4 are mosquito-borne human pathogens that cause widespread epidemics withconsiderable morbidity and mortality. The aim of this study was to evaluate the dengue serotypes profile, which were circulating inSurabaya. This research has been carried out consisting of 360 samples from patients with dengue virus infections, according the WorldHealth Organization (WHO) criteria. These sera were collected from patients Dr. Soetomo Hospital and private laboratory in Surabayafrom 2008–2009. From 360 samples, 68 samples (18.9%) were undifferentiated fever, 53 samples (14.7%) were dengue fever, 239samples (66.4%) were dengue hemorrhagic fever. From 58 Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) samples, 25samples (43%) were positive, consisting of 52% DEN-2, 20% DEN-1, 16% DEN-3 and 12% DEN-4. These results showed that fourserotypes are circulating in Indonesia, dominated by DEN-2, followed by DEN-1, DEN-3 and DEN-4.

scholarly journals RNA ISOLATION OF DENGUE VIRUS TYPE 1 WITH DIFFERENT PRECIPITATION SOLVENTS: DIMETHYL SULFOXIDE, ACETONE, AND ETHANOL 70%

2018 ◽  
Vol 7 (3) ◽  
pp. 62
Author(s):  
Anisa Maharani ◽  
Teguh Hari Sucipto ◽  
Harsasi Setyawati ◽  
Siti Churrotin ◽  
Ilham Harlan Amarullah ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Dimethyl Sulfoxide ◽  
Virus Type ◽  
Dengue Hemorrhagic Fever ◽  
Rna Isolation ◽  
Hemorrhagic Fever ◽  
Chain Reaction ◽  
Polymerase Chain

Dengue Hemorrhagic Fever (DHF) is caused by dengue viruses that belong to Flaviviridae. The disease is known to be caused by 4 types of dengue viruses, namely DENV-1, DENV-2, DENV-3, and DENV-4 associated with antigenic. Dengue virus is a virus RNA that causes illness with clinical manifestations of Dengue Fever, Dengue Hemorrhagic Fever and Dengue Shock Syndrome. The aim of research was to determine the effectiveness of dimethyl sulfoxide, acetone, and ethanol 70% as precipitation solvent in the process of RNA isolation. The method used was Reverse Transcription - Polymerase Chain Reaction (RT-PCR) and Polymerase Chain Reaction (PCR) with specific primers for dengue virus type 1 (DENV-1). RNA isolation can be done easily using an RNA Isolation Kit. Use of RNA Isolation Kit results in a purer RNA isolate from contaminants and from RNA degradation. In generally the isolation is using cold ethanol / alcohol with concentration 90-95%. Ethanol / Alcohol does not dissolve RNA and light density of alcohol lighter than water makes RNA rise and hover on the surface. In RNA isolation solvent precipitation that used are acetone, ethanol 70%, and DMSO. In qualitative RNA measurements using agarose gel electrophoresis and was examined under the UV light-illuminator and quantitative RNA measurements using Nanodrop spectrophotometry with absorbance ratio at 260/280 and 260/230 showed a good result indicated by the appearance of the band on electrophoresis results in PCR. While the measurement quantitatively is showed that there was still protein contamination but the results are quite good because it does not much different from the ratio set in the reference. Acetone, ethanol 70%, and DMSO can be used as a substitute of 96% ethanol in the process of RNA isolation in DENV-1 virus and can also be applied to other dengue virus because the structure of the 4th antigen serotype is very similar one with the other and no effect.

scholarly journals Diversity of Dengue Virus Serotype in Endemic Region of South Sulawesi Province

2018 ◽  
Vol 2018 ◽  
pp. 1-4 ◽  
Author(s):  
Muh. Taslim ◽  
A. A. Arsunan ◽  
Hasanuddin Ishak ◽  
Sudirman Nasir ◽  
Andi Nilawati Usman
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Statistical Test ◽  
Test Results ◽  
Rt Pcr ◽  
Chain Reaction ◽  
South Sulawesi ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Endemic Areas

The objective of this research was to investigate serotype diversity pattern of dengue hemorrhagic fever virus by using real-time-polymerase chain reaction (RT-PCR) method. It was an explorative laboratory research in endemic dengue fever area in South Sulawesi Province, Indonesia, that is, Makassar municipality and Maros and Gowa region. Serological examination was carried out using real-time-polymerase chain reaction (RT-PCR) method to determine the serotype of dengue virus. The data showed that, of 30 patients, 20 patients (66.67%) were from Makassar municipality: 10 patients (33.33%) from Gowa region and 10 patients (33.33%) from Maros region. The serotypes found were DENV-2 and DENV-4 and no DENV-1 and DENV-3 serotypes were found. Makassar municipality and Gowa region have higher infection with serotype DENV-2, that is, 40% of cases compared with Maros, which is 20.0%. Statistical test results showed no significant differences between the three endemic areas. Maros region has the highest infection with serotype DENV-4, that is, 40% of cases compared with Makassar municipality (5.0%) and Gowa region (0%). Statistical test results showed significant differences between the three endemic areas. This result revealed that serotypes obtained in endemic areas of dengue fever in South Sulawesi are DENV-2 and DENV-4 and not serotypes DENV-1 and DENV-3. Makassar municipality has DENV-2 and DENV-4 serotype, infection dominated by DENV-2, while Maros region also has DENV-2 and DENV-4, but DENV-4 is the dominant serotype. Gowa municipality only has DENV-2 serotype infection.

scholarly journals Detection of Pork Gelatin in Jelly Candy Using Fourier Transform Infrared (FTIR) and Polymerase Chain Reaction (PCR)

2021 ◽  
Vol 328 ◽  
pp. 01006
Author(s):  
Jariyah ◽  
Ratna Yulistiani ◽  
Shelma Wharda Afdilah ◽  
Kusuma Wardhani Mas’udah
Keyword(s):  
Polymerase Chain Reaction ◽  
Fourier Transform ◽  
Principal Component ◽  
Dna Amplification ◽  
Fourier Transform Infrared ◽  
Survey Method ◽  
Chain Reaction ◽  
Detection Analysis ◽  
Pcr Method ◽  
Polymerase Chain

This study aims to identify the content of pork gelatin in jelly candy using Fourier Transform Infrared (FTIR) and Polymerase Chain Reaction (PCR) methods. This method provide information to the public in choosing halal and tested food products. By uses a stepwise cluster survey method to obtain a sample then the samples obtained were isolated in gelatin, analyzed using the FTIR spectrophotometer method, and continued with data analysis using PCA (Principal Component Analysis). In addition, DNA detection analysis of pork gelatin was carried out using the PCR method. The results of the study were FTIR spectrum at wavelengths of 1450 – 1300 cm-1, 1543 cm-1, and 2800-3000 cm-1. The classification of gelatin sources in jelly candy with PCA resulted in the proportion value of Principal Component 1 (PC 1) of 39%, the value of the proportion of Principal Component 2 (PC 2) of 31%, the value of the proportion of Principal Component 3 (PC 3) of 14.5% and the cumulative value of PC 1, PC 2, and PC 3 is 84.5%. DNA amplification of jelly candy samples by PCR proved that all jelly candy samples A, B, C, D, and E did not contain pork.

scholarly journals Polymerase chain reaction (PCR) for the rapid detection of Salmonella in desiccated coconut (DC)

CORD ◽  
2007 ◽  
Vol 23 (2) ◽  
pp. 7
Author(s):  
Jayaratne, D.L.
Keyword(s):  
Polymerase Chain Reaction ◽  
Genomic Dna ◽  
Chain Reaction ◽  
Specific Primers ◽  
Routine Testing ◽  
Cultural Methods ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Low Levels ◽  
M1 Type

This study was carried out to develop a polymerase chain reaction (PCR) protocol for the detection of Salmonella in desiccated coconut (DC). For the PCR detection of Salmonella, genomic DNA was extracted using the ‘boiling lyses’ method and the reaction was carried out with Salmonella genus specific primers enabled to amplify 457bp sequence covering invA and invE genes. Samples of DC produced in mills already tested for Salmonella using conventional cultural methods gave identical results with the present PCR method indicating its suitability for adoption in routine testing.  The sensitivity checked using DNA extracted from artificially inoculated DC with serially diluted inoculum of Salmonella M1 type showed that the developed PCR method can be used to detect very low levels of contamination of Salmonella as low as 4 CFU/g in DC.  The method described here reduces the testing and detection time from 6 days to 24 hours ensuring exporters to obtain Salmonella test reports just prior to shipment. 

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