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Journal : Universa Medicina

Multiplex nested polymerase chain reaction for Treponema pallidum using blood is more sensitive than using serum Effendi, Ida; Rosana, Yeva; Yasmon, Andi; Indriatmi, Wresti
Universa Medicina Vol 37, No 1 (2018)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2018.v37.75-84

Abstract

BackgroundSyphilis is a multistage disease transmitted primarily through sexual intercourse. Nowadays, the polymerase chain reaction (PCR) test for Treponema pallidum has been widely used and is expected to overcome problems in diagnostic tests for syphilis. The Treponema pallidum PCR is influenced by type of specimens, PCR methods and target genes. This study aimed to assess the use of blood and serum in multiplex nested PCR for Treponema pallidum, targeting the 23S rRNA.MethodsA cross-sectional study was conducted from April 2015 - April 2016. Sampling was carried out consecutively among patients with clinical features of secondary syphilis who came to Sexually Transmitted Disease (STD) clinics in Jakarta. All sera were also tested with Rapid Plasma Reagin (RPR) and Treponema pallidum Hemagglutination Assay (TPHA) assay, which was considered as the gold standard for this study. We determined the sensitivity and specificity of the multiplex nested PCR for Treponema pallidum using blood and serum.ResultsPCR test was performed on 122 clinical specimens (61 blood and 61 serum). The positive results of PCR test on blood was 22.95% and serum was 6.56%, while the positive results of serology was 68.85%. The sensitivity of Treponema pallidum multiplex nested PCR on blood was 30.95% compared to serum 9.52% (p=0.006). PCR test on blood is able to detect 3.25 times higher than serum. ConclusionThe use of blood has a higher proportion of positives compared to serum in Treponema pallidum multiplex nested PCR using 23S rRNA target gene.
Microscopic examination using negative staining for rapid diagnosis of syphilis Rosana, Yeva; Effendi, Ida; Indriatmi, Wresti; Yasmon, Andi
Universa Medicina Vol. 41 No. 1 (2022)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2022.v41.64-70

Abstract

BACKGROUNDSyphilis is a global health problem, especially in developing countries including Indonesia. Treponema pallidum, the etiologic agent of syphilis, cannot be cultured in vitro. Syphilis has several clinical manifestations, making laboratory testing a very important aspect of diagnosis. Microscopic examination may support the diagnosis but is rarely used in Indonesia. The aim of this study was to evaluate negative staining using the light microscope to detect T. pallidum in syphilitic lesions. METHODSA cross-sectional study was conducted involving 27 subjects who came to several dermato-venereology clinics in Jakarta. Exudates were collected from genital ulcers, condylomata lata, and dry mucocutaneous rash on palms and soles of syphilis patients. Negative staining using one drop of Indian ink was used to examine for treponemas under the light microscope at 10x100 magnification. RESULTSMicroscopic examination using negative staining showed a few clusters of small and spiral shaped bacteria. Of the 39 specimens from 27 subjects, microscopic examinations were successfully done on 10 specimens. Observations could only be conducted on 5 specimens, 3 (60.0%) of which showed the morphology of spirochetes. This examination is the easiest method for detecting the bacteria. Moreover, the bacteria that were isolated from painless genital ulcers could be observed more clearly than those from erythematous maculopapular lesions. CONCLUSIONTreponema pallidum was successfully detected by microscopic examination in all moist lesions, but was difficult to detect in dry lesions. Negative staining under the light microscope appears to be simple, affordable, and available in most microbiology laboratories in Indonesia.
Resistance genes of Neisseria gonorrhoeae to cefixime and azithromycin Louisa Ivana Utami; Yeva Rosana
Universa Medicina Vol. 42 No. 1 (2023)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2023.v42.108-118

Abstract

Gonorrhea is the second most common sexually transmitted bacterial infection (STI), following Chlamydia. Neisseria gonorrhoeae resistant to antibiotics are increasing globally in the world. In recent years, many studies have reported reduced susceptibility of N.gonorrhoeae to almost all clinically useful antibiotics and also reported cases of multi-resistance. Resistance mechanisms for N. gonorrhoeae can occur through genetic and non-genetic changes. Resistance to cefixime and azithromycin as first-line antibiotics for monotherapy recommended by the World Health Organization (WHO) has been reported from several countries. Genetic changes were reported as the main cause of N.gonorrhoeae resistance to cefixime and azithromycin. Based on the WHO and the United States Centers for Disease Control and Prevention recommendations, countries are increasingly using a combination of cephalosporin and azithromycin for the treatment of gonorrhea. The aim of this review is to analyze genetic variation of N.gonorrhoeae resistance to cefixime and azithromycin. Articles published in English in the last 12 years (from 2010 to 2021) were retrieved from Science Direct, PubMed, Springerlink, Oxford and Nature using relevant searching terms. Mutants of cefixime-resistant N.gonorrhoeae are mediated by mosaic and non-mosaic penA genes encoding penicillin binding protein 2. In addition, mutations in the repressor and promoter genes of mtrR were also found that caused overexpression of the microbial efflux pump. Meanwhile, N. gonorrhoeae resistance to azithromycin reportedly occurs through two strategies, namely overexpression of the efflux pump (mutation of the mtrR codon region) and decreased affinity for antibiotics (single base mutation in the 23S rRNA gene). With the limited choice of antibiotics for the management of N.gonorrhoeae, it is necessary to do regular surveillance for monitoring drug resistance. By understanding the mechanism of resistance, the use of these antibiotics can be rationally optimized.
Profile of multidrug-resistant bacteria causing urinary tract infections in inpatients and outpatients in Jakarta and Tangerang Rosana, Yeva; Herliyana, Lina; Krisandi, Grady; Anggraini Suwarsono, Erike
Universa Medicina Vol. 42 No. 3 (2023)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2023.v42.303-313

Abstract

Background Urinary tract infections (UTIs) caused by bacteria occupy the second highest rank of common infectious diseases in the world. Empirical use of antibiotics may give rise to multidrug-resistant (MDR) bacteria because of irrational prescription. Choice of antibiotics to treat UTIs is limited because of MDR bacteria. Thus, this study was conducted to investigate the bacterial antibiotic susceptibility patterns in inpatients and outpatients in Jakarta and Tangerang. Methods Bacterial isolates were obtained from midstream urine specimens from 43 inpatients and 43 outpatients with UTIs in Jakarta and Tangerang. Bacteria were isolated on blood and MacConkey agar media using colony count method. Isolate identification and their susceptibility patterns were performed using VITEK2 compact system according to manufacturers’ instructions. Data were analyzed using Chi-square test. Results A total of 89 bacterial isolates consisting of 15 bacterial species were successfully isolated from 86 specimens. Gram-negative bacteria were the most common etiology of UTIs in inpatients and outpatients. MDR bacteria were found in 52 of the 89 isolates. ESBL-producing E. coli was the most common MDR bacteria. ESBL-producing E. coli and other MDR bacteria showed good susceptibility to ertapenem, meropenem, amikacin, and tigecycline. There were no significant differences regarding the MDR bacterial count in inpatients and outpatients (p=0.521). Conclusion E. coli was found to be the most common MDR bacteria causing UTIs in inpatients and outpatients in Jakarta and Tangerang. Higher resistance to many antibiotics was found in MDR bacterial isolates in inpatients compared to outpatients. MDR bacteria in outpatient UTIs were highly resistant to commonly used antibiotics.