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Hantavirus infection in clinically suspected dengue fever patients Hera Nirwati, Praseno
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 03 (2008)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Background: Hantavirus has been found in many parts of the world, including newly isolated virus in Indonesia. Although infection with the virus can result in specific signs and symptoms known as hemorrhagic fever with renal syndrome (HFRS), clinical presentation of the disease may be similar to those of dengue or other viruses infection. Diagnosis of both dengue and hantavirus infection should be established by laboratory test for the detection of specific antibodies.Objective: The aim of this study was to determine the presence of hantavirus infection in patients suspected to have dengue fever.Methods: Sera were prepared from venous blood of patients. Specific IgG and IgM anti hantavirus in sera from clinically suspected dengue fever patients were examined by an indirect immunoflourescence antibody technique.Results: Eight percent of sera samples were positive for both specific IgG and IgM anti hantavirus, whereas 1 2 percent of samples were positive for IgG only.Conclusion: It is concluded that the recent infection with hantavirus have been found in 8 percent of clinically suspected dengue fever patients, whereas 12 percent of the patients were infected with the virus some time in the past.Key words: hantavirus - immunofluorescence antibody technique - dengue fever - specific IgG and IgM anti hantavirus

The use of bacteriophage therapy for curing the Escherichia coli 0157 infection in mice Metha Restu, Rio Rendy, Hera Nirwati, Susi Iravati, Mova Aria, Ida Ayu Putu
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 03 (2008)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Background: Diarrhea is still a problem in public health, especially in developing country such as Indonesia. Escherichia coli 0157:H7 {Enterohemorrhagic Escherichia coli (EHEC)} is one of the important strains as the etiology of bloody diarrhea with sistemic complications such as hemolytic uremic syndrome and hemorrhagic colitis. The increase in the discovery of E. coli 0157:H7 resistance to antibiotic is a worldwide problem that must be solved. Bacteriophage application can be a promising alternative therapy. In addition, bacteriophage can also be used as diagnostic tool for bacterial identification and as biocontrol agent in bacterial water pollution.ObJective: The aim of this study was to isolate and identify a specific bacteriophage using a specific strain of E. coli 0157 and use this bacteriophage to cure the E. coli 0157 infection in mice.Methods: E. coli was isolated and identified from faecal samples of diarrheal patients from many Primary Health Centers in Yogyakarta, using McConkey Agar and biochemical media. E. coli 0157 was determined using sorbitol McConkey Agar and agglutination test. Toxins of these strains were detected using hemolysis assay method. Bacteriophage was isolated using one of E. coli 0157 strain (E. coli K-151) from river water of Kali Mambu. The therapeutic effect of this bacteriophage was studied using eighteen threemonth-old male mice of Swiss strain. They were classified randomly into three groups (6 groups). Mice in group A and B were infected with 0.5 ml of 108 CFU of E. coli suspension orally. Only mice in group A were treated with bacteriophage 1.10"pfu/mL, while those in group B were not treated, and group C was used as control.Results: Three strains of E. coli 0157 (K-151, K-840 and K-854) were isolated among 70 E. coli isolates. Bacteriophage K-151 was isolated from the river water. Average cure duration in the group who was given phage K-151 therapy was 34.17 hours, and average cure duration in the group that was not given phage K-151 therapy was 72.7 hours. The mortality rate of group A was 0%, while in group B was 17%. Conclusion: Bacteriophage is effective as alternative therapy against E. coli infection in mice.Keywords: E. coli 0157 - bacteriophage - hemolysis assay - diarrhea

Genotyping of Rotavirus by Using RT-PCR Methods Nirwati, Hera; Wibawa, Tri; Aman, Abu Tholib; Soenarto, Yati
Indonesian Journal of Biotechnology Vol 18, No 1 (2013)
Publisher : Universitas Gadjah Mada

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There is a great diversity of rotavirus genotypes circulating worldwide, with dominant genotypes changing from year to year. Rotavirus genotyping was performed by using reverse transcription PCR with type-specifi c-primers. Since rotavirus is a RNA virus that has high mutation rate, there was a possibility of technical diffi culty in genotyping due to mutation in the primer binding sites. During Indonesian rotavirus surveillance study 2006-2009, it was reported that 17% of samples subjected for G type and 21% of samplessubjected for P type were untypeable. The objective of this study was to identify genotypes of the samples that were untypeable previously using RT-PCR based on the method described by Das et al. (1994) and Gentsch et al. (1992). There were 30 samples subjected to G type and 61 samples subjected to P type to be re-typed using method described by Gouvea et al. (1990) and Simmond et al. (2008) for G and P typing, respectively. By using another set of primer, the genotype of all samples was identifi ed. This study highlights the importance of a constant reconsideration of primer sequences employed for the molecular typing of rotaviruses.Key words: rotavirus, G typing, P typing

An increased susceptibility of Staphylococcus aureus to tetracyclin during exposure to rifampicin Hera Nirwati, Hera Nirwati
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 30, No 03 (1998)
Publisher : Universitas Gadjah Mada

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acterial resistance to antibiotics has been an important problem worldwide. Discoveries and clinical use of new antibiotics have always been followed by emergence of strains of bacterial resistant to the antibiotics in question within relatively short period of time. Various efforts have been attempted to overcome the problem, one of which being the use of two or more different antibiotics in combination. In this study the effect of rifampicin on alternation of susceptibility of S. aureus to tetracycline was evaluated. Sixty seven isolates of tetracycline resistant S. aureus determined by standard disk diffusion method of Kirby-Bauer were collected during the period from August 1997 to January 1998. Minimal Inhibitory Concentration (MIC = KHM) values of rifampicin for this isolates were determined by broth dilution method. MIC of tetracycline for each isolate was determined while the bacteria were exposed to rifampicin at sub MIC. At the same time determination of the MIC was also performed without exposing the bacteria to rifampicin and this procedure served as control. Results showed that 91% (61 of 67) rifampicin - exposed isolates have increase susceptibility to tetracycline as reflected by the decrease of MIC values. It is concluded that there is synergistic effects of tetracycline and rifampicin on S. aureus. The number of isolate with increased susceptibility to tetracycline during exposure to rifampicin was statistically significant (p < 0.05).Key Words : rifampicin - Staphylococcus aureus - susceptibility - tetracycline

Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR Wijayanti, Nastiti; Wibawa, Tri; Nirwati, Hera; Haryanto, Aris; S, Sutaryo1
Indonesian Journal of Biotechnology Vol 11, No 2 (2006)
Publisher : Universitas Gadjah Mada

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world. We have evaluated the combination of one-step RT-PCR and multiplex nested PCR assays for detectingdengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primersthat conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirusand there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virususing multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assaycan be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiologyand also evolutionary studies.Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR

Utilization of Closed Circuit Television in Improving Nurse's Compliance on Hand Hygiene in Budhi Asih Hospital Jakarta Ramadhanti, Ahijrah; Dwiprahasto, Iwan; Nirwati, Hera
Jurnal Keperawatan Soedirman Vol 15, No 2 (2020)
Publisher : Jurusan Keperawatan FIKES UNSOED

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.jks.2020.15.2.1204

Health-care associated infections (HAIs)Â are infectionsoccurringÂ inhospitalized patients.The most effective way to preventÂ HAIsisÂ throughhand hygiene. However, hand hygiene compliance in health workers is still low.Â This research aimed toÂ understandtheassociation between CCTVutilizationasÂ aÂ reminder toolÂ inimproving the nurses'hand hygiene compliancein Budhi AsihHospitalJakarta. The studyÂ used aÂ quantitativemethod bya quasi-experimentalapproach. The 60 subjectsÂ weredivided into two groups:Treatment and Control Groups based on their workplace. Quantitative data wereobtained by filling-in a WHO-standardized questionnaire and observing each group before and after an intervention.Â Data wereÂ analyzed by univariate and bivariate analyses with chi-square test and multivariate analysis with logisticregression test.Â Nurses' hand hygiene compliance through CCTV observation in Budhi Asih Hospital was 57%. The use of CCTV as reminder media significantly improved hand hygiene compliance (p = 0.002), compliance to 6 steps (p = 0.002) and compliance to the standard time of hand hygiene (p = 0.003). There was no significant correlation between individual characteristics (sex, age, education, working experience, and infection control training participation) with nurses' compliance on hand hygiene.Â The use of CCTV as reminder media significantly improved nurses' compliance to do hand hygiene.Keywords:Â CCTV, Reminder, Hand Hygiene, Complience.Â

IDENTIFICATION OF THE VIRUS DENGUE–3 EPITOPE’S IMMUNODOMINAN USING THE SYNTHETIC PEPTIDE Nirwati, Hera; ., Sutaryo; Wahyono, Djoko
Indonesian Journal of Pharmacy Vol 13 No 1, 2002
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

Dengue virus infection has been known as an important health problem in many tropical countries, because of increasing number of patients, expansion of epidemic areas and emergence of severe clinical manifestations. Dengue virus consists of 3 structural and 7 nonstructural proteins. The major virion surface protein, the envelope protein, is the most important antigen with regards to virus biology and humoral immunity. Synthetic peptide derived from the envelope protein gene sequence can be used to identify region of the envelope protein that elicits antibodies. One hundred and sixty-one synthetic peptides were synthesized based on sequence of protein envelope dengue-3 virus published by Osatomi and Sumiyoshi. Each peptide consists of 15 amino acids with the last 12 amino acids overlapped. Peptide synthesizer, auto-spot robot ASP 222 (Abimed), was used for synthesis using solid spot method developed by Ronald Frank. Synthetic peptides attached to nitrocellulose membrane were used for spot immunoassay of 6 normal human sera, 6 dengue infected human sera and 6 non dengue-infected. Nine peptides were specific for dengue infected sera and these might be candidate immunodominat epitopes of dengue virus-3. They were EGLSGATWVDVVLEH (amino acids number 13-27), VCKHTYVDRGWGNGC (91-105),SIEGKVVQHENLKYT (124-138), TVHTGDQHQVGNETQ (142-156), TLGLECSPPTGLDFN (178-192), KGEDAPCKIPFSTED (325-339), PFSTEDGQGKAHNGR (334-348), GARRMAILGDTAWDF (406-420) and KIGIGVLLTWIGLNS (454-468). Based on spot immunoassay, 11 peptides were synthesized and used them for ELISA of 15 normal sera, 22 non-dengue infected sera and 37 dengue sera. Peptides BTLDDIELQKTEATQLA, BPFSTEDGOGKAHNGR and BKKEEPV NIEAEPPFG showed significantly different in their reactivities to the three groups. Combinations of two peptides were used for ELISA. Only one of them (BKGEDAPOKIPFSPED and BRMAILGD TAWDFGSV) showed significantly different in their reactivi- ties to the three groupsKey words: synthetic peptide, envelope protein dengue-3, spot immunoassay

Rapid Detection and Molecular Typing of Dengue Virus by Using Multiplex-Nested-RT-PCR Nastiti Wijayanti; Hera Nirwati; Tri Wibawa; Aris Haryanto; S. Sutaryo
Indonesian Journal of Biotechnology Vol 11, No 2 (2006)
Publisher : Universitas Gadjah Mada

world. We have evaluated the combination of one-step RT-PCR and multiplex nested PCR assays for detectingdengue viruses from clinical samples. Twelve patients were screened for the dengue virus, using a pair of primersthat conserve for several Flavivirus. The results showed that in 12 suspect patients, 100% were positive for Flavivirusand there are some genotypic variation among them, that indicated by several RT-PCR products higher than 511 bp,the expected product for RT-PCR. Further assay was performed to clarify the presence and serotypes of dengue virususing multiplex nested PCR. Serotyping results indicated that 83,3% of samples can be confirmed for dengue virus.Among the dengue virus positive 16,7 % are dengue-2, 16.7 % are dengue-3, and the most common 50% are dengue-4,whereas dengue-1 were not found among the patients. The combination of RT-PCR and multiplex nested PCR assaycan be used for rapid analysis dengue samples in early phase which is potentially useful for clinical, epidemiologyand also evolutionary studies.Key words: Flavivirus, dengue virus, serotype, RT-PCR, multiplex nested PCR

The use of bacteriophage therapy for curing the Escherichia coli 0157 infection in mice Hera Nirwati, Susi Iravati, Mova Aria, Ida Ayu Putu Metha Restu, Rio Rendy
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 40, No 03 (2008)
Publisher : Journal of the Medical Sciences (Berkala Ilmu Kedokteran)

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Background: Diarrhea is still a problem in public health, especially in developing country such as Indonesia. Escherichia coli 0157:H7 {Enterohemorrhagic Escherichia coli (EHEC)} is one of the important strains as the etiology of bloody diarrhea with sistemic complications such as hemolytic uremic syndrome and hemorrhagic colitis. The increase in the discovery of E. coli 0157:H7 resistance to antibiotic is a worldwide problem that must be solved. Bacteriophage application can be a promising alternative therapy. In addition, bacteriophage can also be used as diagnostic tool for bacterial identification and as biocontrol agent in bacterial water pollution.ObJective: The aim of this study was to isolate and identify a specific bacteriophage using a specific strain of E. coli 0157 and use this bacteriophage to cure the E. coli 0157 infection in mice.Methods: E. coli was isolated and identified from faecal samples of diarrheal patients from many Primary Health Centers in Yogyakarta, using McConkey Agar and biochemical media. E. coli 0157 was determined using sorbitol McConkey Agar and agglutination test. Toxins of these strains were detected using hemolysis assay method. Bacteriophage was isolated using one of E. coli 0157 strain (E. coli K-151) from river water of Kali Mambu. The therapeutic effect of this bacteriophage was studied using eighteen threemonth-old male mice of Swiss strain. They were classified randomly into three groups (6 groups). Mice in group A and B were infected with 0.5 ml of 108 CFU of E. coli suspension orally. Only mice in group A were treated with bacteriophage 1.10"pfu/mL, while those in group B were not treated, and group C was used as control.Results: Three strains of E. coli 0157 (K-151, K-840 and K-854) were isolated among 70 E. coli isolates. Bacteriophage K-151 was isolated from the river water. Average cure duration in the group who was given phage K-151 therapy was 34.17 hours, and average cure duration in the group that was not given phage K-151 therapy was 72.7 hours. The mortality rate of group A was 0%, while in group B was 17%. Conclusion: Bacteriophage is effective as alternative therapy against E. coli infection in mice.Keywords: E. coli 0157 - bacteriophage - hemolysis assay - diarrhea

Potential secondary metabolite analysis of soil Streptomyces sp. GMR22 and antibacterial assay on Porphyromonas gingivalis ATCC 33277 Hera Nirwati; Ema Damayanti; Eti Nurwening Sholikhah; . Mustofa; Jaka Widada
Journal of the Medical Sciences (Berkala Ilmu Kedokteran) Vol 54, No 2 (2022)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.19106/JMedSci005402202202

Infectious diseases caused by oral pathogenic bacteria are currently a serious problem due to the increasing incidence of antimicrobial resistance. Streptomyces sp. GMR22, a soil actinobacterium which has large-genome size. In previous studies, it was known to have antifungal, and antibiofilm activity on Candida albicans. However, its antibacterial activity on oral pathogenic bacterium, Porphyromonas gingivalis is not clear. This study aimed to identify potential active compound based on genome mining analysis and to evaluate the antibacterial activity of GMR22 extract on P. gingivalis ATCC 33277. Potential active compounds and biosynthesis gene clusters were analysis using antiSMASH version 5. Antibacterial activity assay was carried out by the microdilution method on P. gingivalis ATCC 33277. Based on genome mining analysis polyketide synthase (PKS), the Streptomyces sp. GMR22 is the abundant BGCs (35%) and has large-predicted compounds which have antibiotic-antibacterial activity (22.9%). On antibacterial assay, chloroform extract of GMR22 at 7.8 – 62.5 µg/mL has high antibacterial activity on P. gingivalis compared to other extracts. Soil Streptomyces sp. GMR22 bacterium has biotechnological potential to produce active compounds for antibacterial.

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