Diagnostic value of the QuickStripe™ chlamydia rapid test among high-risk women in Jakarta.

BACKGROUND: Difficulty in diagnosing Chlamydia trachomatis infections, including chlamydial cervicitis, is a notable challenge in managing sexually transmitted infections in Indonesia. Gram staining is usually done to make a presumptive diagnosis despite its low sensitivity and specificity. Polymerase chain reaction (PCR) is considered the gold standard, but it is costly, technically demanding, and difficult to be performed in low-resource settings. Thus, rapid point-of-care tests with high sensitivity and specificity are needed to diagnose chlamydial cervicitis. METHODS: This cross-sectional study included symptomatic and asymptomatic high-risk women in the Mulya Jaya Sex Workers Rehabilitation Center in June to July 2020. Endocervical swabs from each participant were taken for QuickStripe™ chlamydia rapid test (CRT), Gram staining, and real-time PCR. RESULTS: A total of 41 participants were enrolled. The sensitivity and specificity for QuickStripe™ CRT were 73.6% (95% CI: 48.80%-90.85%) and 81.82% (95% CI: 59.72%-94.81%). Positive and negative predictive values were 77.78% (95% CI: 58.09%-89.84%) and 78.05% (95% CI: 62.39%-89.44%). Proportion of chlamydial cervicitis in study participants based on real-time PCR was 46.3%. CONCLUSIONS: We concluded that QuickStripe™ CRT can be recommended as an alternative diagnostic test for high-risk populations in Jakarta.

Detection of A2058G and A2059G on the 23S rRNA Gene by Multiplex Nested PCR to Identify Treponema pallidum Resistance to Azithromycin in Indonesia.

Azithromycin is an antibiotic used to treat syphilis, especially in the context of penicillin allergy. Although resistance to azithromycin has been widely reported to be associated with one- and/or two-point mutations on the 23S rRNA gene, it has yet to be described in Indonesia. Specimens were collected from 220 patients diagnosed with secondary syphilis. A multiplex nested polymerase chain reaction (PCR) testing system using the 23S rRNA target gene of Treponema pallidum was designed using three primer pairs. The first step involved the use of PCR primer pairs to detect T. pallidum. In the second step, two PCR primer pairs were constructed to identify azithromycin-resistant T. pallidum based on A2058G and A2059G point mutations. T. pallidum detected in samples from Jakarta or Bandung were not resistant to azithromycin. However, azithromycin-resistant T. pallidum were found in samples from Makassar, Medan, and Bali. Specimens from heterosexual males and patients with HIV accounted for the majority of azithromycin resistance noted in the study. This study demonstrated that the azithromycin-resistant T. pallidum detected in Indonesia appear to be a novel variant of resistance, containing both the A2058G and A2059G mutations found in Medan and Makassar.

Genetic characterization of diphtheria tox B to evaluate vaccine efficacy in Indonesia.

Background and Objectives: Blocking the attachment of diphtheria toxins to host cells through the intact receptor binding site (tox B) was the initial mechanism of action of the diphtheria vaccine. Diphtheria outbreaks in populations with good vaccination coverage can be caused by mutations or changes in the genetic structure of the tox B protein. The aim of this study was to characterize the Tox B protein produced by Corynebacterium diphtheriae isolated from 2018 to 2019 in patients in Jakarta who had already received the diphtheria vaccine. Materials and Methods: Of the 89 throat swab specimens of patients with a clinical diagnosis of diphtheria, 10 were positive for diphtheria and toxin. PCR was used to amplify the tox B DNA fragment in the 10 positive isolates. DNA sequencing was conducted with overlapping primers and the DNA sequences were analysed by using SeqScape V2.7. Results: Of the 10 isolates, nine isolate showed a DNA mutation (G30A), but the mutation did not change the amino acid encoding arginin (silent mutation). Our findings indicate that the efficacy of the diphtheria vaccine used in Indonesia has not decreased because of mutations in the tox B genes not change the amino acid. Conclusion: Overall, there are no amino acid changes in the tox B protein, indicating that the outbreaks are not affected by mutation in tox B. Another possible mechanism - overexpression of the toxin - is likely responsible for causing diphtheria in patients who have a complete history of immunization in Indonesia.

Urinary Tract Infections among Indonesian Pregnant Women and Its Susceptibility Pattern.

Pregnant women are usually at risk of urinary tract infections (UTIs) such as asymptomatic bacteriuria. In the current multidrug-resistance era, appropriate diagnosis and treatment should be provided to avoid complications in pregnant women in developing countries, which have limited facilities, such as Indonesia. The aim of this study was to evaluate in vitro susceptibility tests. Urinary isolates were collected from 715 pregnant women who visited eight Community Health Centers in Jakarta, Indonesia, between 2015 and 2017. We identified bacterial uropathogens from samples that were positive for nitrite/leukocyte esterase (LE), using two types of VITEK cards. Since noncompliance among patients is a major problem, fosfomycin-trometamol 3 g single-dose sachets were given to the patients, and the side effects of the medication and neonatal outcomes were reported. Asymptomatic bacteriuria was found in 10.5% of the 715 pregnant women. Escherichia coli was the most common etiological factor (26.7%), followed by Klebsiella pneumoniae (20%), Streptococcus agalactiae (9.3%), Enterobacter cloacae (5.3%), Enterococcus faecalis (5.3%), Staphylococcus saprophyticus (4%), Acinetobacter baumannii (4%), and others. Out of 76 pregnant women who took fosfomycin-trometamol, two complained of diarrhea that subsided without medication and fever that responded to paracetamol. Neonatal outcomes showed 100% full-term and normal-weight babies. E. coli, including extended-spectrum beta-lactamase- (ESBL-) producing E. coli, was 100% susceptible to fosfomycin. Nitrite/LE test results are often used as evidence for empiric antibiotic administration for treating asymptomatic bacteriuria in pregnancy, but the diagnosis should be confirmed using culture tests. Based on in vitro susceptibility patterns and medication outcomes, fosfomycin-trometamol single dose could be administered to noncompliant UTI patients, including pregnant women.

Detection of toxin-producing Corynebacterium diphtheriae from throat swabs of diphtheria patients using duplex real-time PCR.

BACKGROUND AND OBJECTIVES: Diphtheria is a potentially fatal disease caused by toxigenic bacterial infection, particularly from Corynebacterium diphtheriae (C. diphtheriae). Isolation of C. diphtheriae is technically lacking in sensitivity, and Elek's test to detect toxin production has several difficulties associated with its application. Duplex real-time PCR to throat swab of suspected diphtheria patients can detect both bacteria and toxin-encoding genes simultaneously, faster, with higher sensitivity and specificity. MATERIALS AND METHODS: A total of 89 consecutive throat swabs from suspected diphtheria patients were collected from Sulianti Saroso Infectious Disease Hospital, Jakarta, during 2018 to 2019. Two pairs of primers and probes, targeting the rpoB gene of C. diphtheriae and the A-subunit of the diphtheria toxin gene, were used in this study. Parameters including annealing temperature, concentration of primers and probes, inhibitors, cross-reaction and detection limit were all optimized. Elek's toxigenicity test and clinical data were analyzed for comparison. RESULTS: The optimum annealing temperature was 55°C. The concentrations of Cd primer, Tox primer, Cd probe and Tox probe were 0.4, 0.6, 0.5 and 0.625 µM, respectively. DNA elution and template volumes were 50 µL and 5 µL. The detection limit was 2 CFU/mL. No cross-reaction with other microorganisms was observed. Of the 89 samples, duplex real-time PCR gave better results than the standard test, with 19 (21.3%) and 10 (11.2%) patients diagnosed with diphtheria, respectively. CONCLUSION: Duplex real-time PCR increases the rate of laboratory diagnosis of diphtheria, compared to the standard method to detect potentially toxigenic C. diphtheriae.

Formulation, characterization, and in vitro testing of azelaic acid ethosome-based cream against Propionibacterium acnes for the treatment of acne.

Azelaic acid is an antiacne drug by inhibiting thioredoxin reductase enzyme of Propionibacterium acnes (P. acnes) that affects the inhibition of bacterial DNA synthesis which occurs in the cytoplasm. Azelaic acid must penetrate through the stratum corneum to the sebaceous tissue and into cytoplasm by passing through thick peptidoglycan of P. acnes. Thus, it is necessary to increase the penetration of azelaic acid that formulated based ethosome. This study using thin-layer hydration method forms an ethosomal suspension with variations of concentration ethanol (30%, 35%, and 40%). Antibacterial activity was conducted using broth dilution method to determine minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The antibacterial activity of azelaic acid ethosome cream based was compared with the marketed cream (Zelface® cream). Azelaic acid ethosome with 35% ethanol has given best result with entrapment efficiency of 94.48% ± 0.14%. Antibacterial activity to P. acnes showed that azelaic acid ethosome-based cream was given better activity than marketed cream (Zelface® cream). The value of MIC and MBC of azelaic acid ethosome-based cream was 250 µg/ml while the marketed cream (Zelface® cream) was shown MIC of 250 µg/ml and MBC of 500 µg/ml. This study proved that the azelaic acid ethosome-based cream has better antibacterial activity.

Overexpression and mutation as a genetic mechanism of fluconazole resistance in Candida albicans isolated from human immunodeficiency virus patients in Indonesia.

Fluconazole is the standard treatment for oropharyngeal candidiasis, which is the third most common opportunistic infection in human immunodeficiency virus (HIV)/AIDS patients in Indonesia. Overuse of this drug could lead to the emergence of resistance. The objective of this study was to analyse the role of ERG11, CDR1, CDR2 and MDR1 gene overexpression and mutations in the ERG11 gene as a genetic mechanism of fluconazole resistance in Candida albicans isolated from HIV patients in Indonesia. Overexpression of ERG11, CDR1, CDR2 and MDR1 was analysed by real-time reverse transcription PCR, while ERG11 gene mutation analysis was performed using sequencing methods. Seventeen isolates out of 92 strains of C. albicans isolated from 108 HIV patients were found to be resistant to azole antifungals. The highest gene overexpression of ERG11 was found in C. albicans resistant to single fluconazole, while the highest gene overexpression of CDR2 was detected in all isolates of C. albicans resistant to multiple azoles. Amino acid substitutions were observed at six positions, i.e. D116E, D153E, I261V, E266D, V437I and V488I. The amino acid substitution I261V was identified in this study and was probably associated with fluconazole resistance. The combination of overexpression of CDR2 and ERG11 and mutation in the ERG11 gene was found to be a genetic mechanism of fluconazole resistance in C. albicans isolated from HIV patients in Indonesia.

Trends in antimicrobial susceptibility of gram-negative bacteria isolated from blood in Jakarta from 2002 to 2008.

BACKGROUND: This study examined the susceptibility of Gram-negative bacteria in the bloodstream to antimicrobials with the aim of providing information relevant to the guidance of therapy. METHODOLOGY: Blood specimens received by the Laboratory of Clinical Microbiology, Faculty of Medicine, University of Indonesia, from 2002 to 2008, were analyzed for the presence of Gram-negative bacteria and their susceptibility to four antibiotic groups frequently administered in hospitals and community settings. RESULTS: During the seven-year period leading up to 2008, approximately 68% of Gram-negative bacteria were identified among all positive isolates from blood specimens. The eight most frequent species found were Acinetobacter anitratus (25.8%), Pseudomonas aeruginosa (19.5%), Klebsiella pneumoniae subsp. pneumoniae (14.5%), Enterobacter aerogenes (8%), Salmonella Typhi (7.5%), Escherichia coli (6.2%), Alcaligenes faecalis (5.6%) and Klebsiella oxytoca (3.2%). At 80% susceptibility or greater, Ceftriaxone and Cefotaxime were active only on E. coli and S. Typhi. Cefepime demonstrated activity on all eight species tested except K. pneumonia while Amikacin showed activity against five species, A. faecalis, E. aerogenes, E. coli, K. pneumoniae subsp. pneumoniae and S. Typhi. Gentamycin was active against three species: E. aerogenes, K. oxytoca and S. Typhi. Ciprofloxacin and Levofloxacin significantly differed in their spectrum: while Ciprofloxacin was active against four of the eight species tested (E. aerogenes, E. coli, K. oxytoca, and S. Typhi ), Levofloxacin was similar to Cefepime and was active against all eight species except K. pneumoniae subsp. pneumonia. CONCLUSIONS: Since antimicrobials are broadly used in Jakarta, it is important that the information captured in this study be disseminated.

Antimicrobial susceptibilities of Neisseria gonorrhoeae strains isolated in Java, Indonesia.

BACKGROUND: Despite the rapid spread of antibiotic resistance in gonococci all over Southeast Asia, there is only limited surveillance for antibiotic susceptibility in Indonesia. GOAL: This study was undertaken to determine the frequency and diversity of antimicrobial resistance in gonococcal isolates from cohorts of female commercial sex workers in Bandung and Jakarta, Indonesia, and to characterize the Tet M plasmid among the tetracycline-resistant strains N gonorrhoeae. STUDY DESIGN: The antimicrobial susceptibility of 267 strains (85 strains from Bandung and 182 from Jakarta) to penicillin, spectinomycin, tetracycline, ciprofloxacin, cefotaxime, thiamphenicol, kanamycin, azithromycin, and trimethoprim-sulfamethoxazole (TMP-SMZ) was determined by agar dilution. Typing of the Tet M plasmid in tetracycline-resistant isolates was performed by PCR. RESULTS: Prevalence of penicillin and tetracycline resistance was extremely high: 60.0% of the isolates from Bandung and 70.9% of the isolates from Jakarta were resistant to penicillin. Of these, 60.0% and 62.1%, respectively, were penicillinase-producing N gonorrhoeae (PPNG). All the isolates from Bandung and 98.4% from Jakarta were resistant to tetracycline. All tetracycline-resistant isolates from Bandung and 97.8% from Jakarta carried a PCR fragment characteristic of the "Dutch" type Tet M plasmid. One isolate from Jakarta showed chromosomal resistance to tetracycline (0.6%). Chromosomal resistance to thiamphenicol (MIC, >/=2.0 microg/ml) was significantly higher in Jakarta than in Bandung (P < 0.05). All gonococcal isolates were susceptible to kanamycin, spectinomycin, cefotaxime, ciprofloxacin, and azithromycin. CONCLUSION: Spectinomycin, fluoroquinolones, and azithromycin are still useful primary drugs for treatment of gonococcal infections in Indonesia. Continued surveillance of antimicrobial susceptibility should be part of gonorrhea control in Indonesia.