Hepatitis C virus RNA detection and HCV genotype in patients with chronic non-A, non-B hepatitis in Jakarta.

Antibody response in HCV infection may be variable and the variability of the serological response could be due to the differences in HCV strains. Since the distribution of hepatitis C virus genotype has been found to be geographically dependent, it is important to determine the distribution of HCV genotype in various countries with high prevalence of chronic non-A, non-B hepatitis. In this study, serum HCV RNA was examined in 53 patients suspected of chronic non-A, non-B hepatitis with an anti-HCV test as determined by currently available assay. HCV viremia was detected in 48 patients (90.6%). These patients had elevated serum ALT level at the time of HCV RNA determination. Using specific genotype probes, all isolates were classified into three different genotypes. Double and triple infections were also noted. HCV genotype 1b is the predominant genotype found in chronic hepatitis C patients in Jakarta.

Persistent, recurring diarrhea in a colony of orangutans (Pongo pygmaeus) caused by multiple strains of Campylobacter spp.

A colony of 10 orangutans (Pongo pygmaeus) experienced persistent, recurring diarrhea caused by multiple infections with Campylobacter jejuni and C. coli. Infections appeared to have occurred through several mechanisms, including fecal-oral transmission between orangutans, and possibly transmission by houseflies contaminated with the organisms from nearby chicken feces. Among the 14 fecal and environmental C. jejuni isolates, 4 different antibiotic susceptibility profiles were detected; there were also 4 different profiles among the 8 isolates of C. coli. In 5 orangutans, there were back-to-back infections by different strains of C. jejuni, suggesting that a single C. jejuni infection may not confer protective immunity against heterologous strains circulating in the same vicinity. Transmission was effectively interrupted by environmental modifications and a 7-day course of oral erythromycin.

Effects of desipramine on rat behavior are prevented by concomitant treatment with ethanol.

Ethanol prevents the decrease of the number of beta-adrenoceptors in the cerebral cortex induced by chronic treatment of rats with desipramine. The activation of the adenylate cyclase, the second messenger, by beta-adrenergic agonists is reduced somewhat less than after treatment with desipramine alone. The present paper examined the hypothesis that ethanol inhibits the neuronal adaptation to desipramine chronic treatment at the functional level as well. Desipramine reduced exploratory behavior (crossings, rearings) as did ethanol. Combined treatment attenuated the effect of desipramine. Cognitive performance was investigated using an active avoidance paradigm. Desipramine-treated rats did not learn the task in contrast to control animals. Again, combination treatment with ethanol improved the ability of the rats to perform the task. The activity of cerebral beta-adrenergic mechanisms was assessed by injection of salbutamol, a beta-adrenoceptor agonist in rats pretreated with 5-hydroxytryptophan (5-HTP). The augmentation of the 5-HTP-induced wet dog shake behavior by salbutamol was observed in all animals independent of the chronic treatment. However, rats treated with desipramine were less active than those treated with tap water or ethanol. The effect of desipramine in the presence of a high concentration of salbutamol was attenuated by ethanol. The observed increase of the number of wet dog shakes correlates with the function of these receptors. In two paradigms, spontaneous motility and apomorphine-induced hypothermia, ethanol did not affect the action of desipramine. It is noteworthy that desipramine acted in both situations within a short time period (minutes to hours). The findings strongly suggest that ethanol can prevent adaptive changes in the brain induced by chronic treatment with the antidepressant desipramine. This is of special interest since the adaptation of beta-adrenoceptors is thought to be critical for the antidepressant efficacy of various therapeutic interventions applied in psychiatric practice.

Simultaneous determination of amoxycillin trihydrate and clavulanate potassium in pharmaceutical preparations by thin-layer chromatography/densitometry.

A simple and rapid densitometric method has been developed for the simultaneous determination of amoxycillin trihydrate and clavulanate potassium in pharmaceutical preparations. After extraction of the analytes with distilled water, the extracts were spotted on precoated silica gel plates, which were then eluted with butyl acetate-methanol-glacial acetic acid-water (15 + 7.5 + 7.5 + 3, v/v). Quantitative evaluation was performed by measuring the absorbance reflectance of the analyte spots at lambda = 240 nm. The densitometric method is rapid, selective, precise, and accurate and, thus, can be used for routine analysis of pharmaceutical preparations in quality control laboratories of the pharmaceutical industry.

Comparative sensitivity of laboratory methods to diagnose dengue virus infections at Husada Hospital, Jakarta.

Several methods are available for diagnosis of dengue virus infections including a new commercially available dengue blot IgG assay. We conducted a study to compare the sensitivity of the dengue blot with the conventional diagnostic methods. Serum samples from suspected dengue patients were collected for virus isolation and the following serological assays: the hemagglutination-inhibition assay, an IgM/IgG enzyme-linked immunosorbent assay, and the dengue blot. When suspected dengue samples were tested by all methods, viral isolation detected the fewest dengue infections (10.5%), while the IgM/IgG ELISA was the most successful (46.3%) in diagnosing dengue infections. In a specific comparison between the IgM/IgG ELISA and the dengue blot, the dengue blot had an overall sensitivity of 48.8%, with a specificity of 88.7%. When patients were classified by their serological response, the dengue blot had a sensitivity of only 1.7% in those patients with a primary or recent dengue infection, however in secondary infections, the sensitivity of the dengue blot improved to 93.5%. Testing convalescent samples from patients with primary infections, only slightly changed the sensitivity of the dengue blot. The diagnosis of dengue is needed rapidly by clinicians to insure prompt treatment of patients. The dengue blot provides a rapid and easily performed assay, especially sensitive in secondary dengue infections which are most common in hospitalized cases in Asia.

Prevalence of hemagglutination-inhibition and neutralizing antibodies to arboviruses in horses of java.

A study was conducted to measure the prevalence of hemagglutination-inhibition (HI) and neutralizing antibodies against two arboviruses (Chikungunya and Japanese encephalitis virus) in horses of Java, Indonesia. Blood specimens were collected from a sample of 112 horses at two stables: Pulo Mas, a racing track-horse complex, located in a residential area in North Jakarta, and Pamulang, a riding school, located in a rural environment of West Jaya. Sera were tested by the HI assay and plaque reduction neutralization test. JEV antibodies were detected by HI in 58 (52%) of the horses, while only 11 (10%) had Chikungunya antibodies by HI. The proportion of Pamulang horses infected with JEV (66%) was significantly higher than found among Pulo Mas horses (40%) screened (p < 0.01). Of the 58 horses with JEV antibodies by HI, 52 (90%) were found to have specific neutralization antibodies to JEV. HI and neutralization tests on horse sera indicated that the risk to alpha virus infections was minimal in horses surveyed from Java. However, there was a high risk of JEV infection among the same population.

Seroprevalence of hepatitis A virus and varicella zoster antibodies in a Javanese community (Yogyakarta, Indonesia).

Hepatitis A virus (HAV) cause an acute inflammation of the liver. Varicella-zoster virus (VZV) cause chickenpox (varicella) and herpes zoster. Effective vaccines against hepatitis A and varicella are available for children, adolescents and adults. In order to implement an appropriate vaccination policy, a baseline to assess the potential benefits and sections of the population who would benefit most are required. We investigated seroprevalence of hepatitis A virus and varicella zoster antibodies in a Javanese community. A total of 1,103 subjects were studied. The 600 subjects aged 4 to 9 years were sampled between 23 October and 2 November, 1995. The other subjects were sampled between 12 October and 1 November, 1996. The overall prevalence of anti-HAV in cohort was 28.7%. Anti-HAV seroprevalence rates were below 30% until the age of 15 and below 40% until the age of 25. The anti-varicella seroprevalence showed only in two thirds of seropositive population at the age of 15. The results of the study have implications for vaccination strategies for both hepatitis A and varicella zoster.

PARK2/PACRG polymorphisms and susceptibility to typhoid and paratyphoid fever.

Host genetic factors may contribute to susceptibility to and outcome in infectious diseases. Recently polymorphisms in PARK2/PACRG, a gene cluster linked to ubiquitination and proteasome-mediated protein degradation, were found to be associated with manifest infection by M. leprae. Here, we address whether these polymorphisms are associated with susceptibility to infection with Salmonella typhi and S. paratyphi A, intracellular pathogens that upon infection of humans share with mycobacteria aspects of the hosts' immune response. The polymorphisms of PARK_e01(-697), PARK2_e01(-2599), rs1333955 and rs1040079 were analysed by polymerase chain reaction and restriction fragment length polymorphism in a case-control study of typhoid and paratyphoid fever patients in an endemic area in Jakarta, Indonesia. For this study, samples were obtained from patients with blood culture-confirmed typhoid fever (n=90), paratyphoid fever (n=26) and fever controls (n=337) in a passive, community-based surveillance and compared to those of randomly selected community controls (n=322) from the same city area. The PARK2_e01(-2599) allele T was significantly associated with typhoid and paratyphoid fever (OR: 1.51, 95%CI: 1.02-2.23) but the other polymorphisms, PARK2_e01(-697), rs1333955 and rs1040079, were not associated. Although within the PARK2/PACRG gene cluster the PARK2_e01(-2599) allele T was most strongly associated with leprosy (OR approximately 3-5), the association with typhoid is much less strong. Our findings suggest that this polymorphism in PARK2/PACRG plays a small but significant role in susceptibility to the intracellular pathogens S. typhi and S. paratyphi.

Helicobacter pylori infection and typhoid fever in Jakarta, Indonesia.

We evaluated the association between typhoid fever and Helicobacter pylori infection, as the latter microorganism may influence gastric acid secretion and consequently increase susceptibility to Salmonella typhi infection. Anti-H. pylori IgG and IgA antibody titres (ELISA) and gastrin concentration (RIA) were determined in the plasma of 87 blood culture-confirmed typhoid fever cases (collected after clinical recovery) and 232 random healthy controls without a history of typhoid fever, in the Jatinegara district, Jakarta. Patients with typhoid fever more often than controls were seropositive for H. pylori IgG (67% vs. 50%, P<0.008), when antibody titres were dichotomized around median titres observed in controls. H. pylori IgA seropositivity was not associated with typhoid fever. Plasma gastrin concentrations indicative of hypochlorhydria (i.e. gastrin > or =25 or > or =100 ng/l) were not significantly elevated in typhoid fever cases compared to controls (P=0.54 and P=0.27 respectively). In a multivariate analysis, typhoid fever was independently associated with young age (<33 years, median age of the controls) [odds ratio (OR) 7.93, 95% confidence interval (CI) 3.90-16.10], and H. pylori IgG seropositivity (OR 1.93, 95% CI 1.10-3.40). Typhoid fever was independently associated with H. pylori IgG seropositivity, but not with elevated gastrin concentration. Therefore, the association suggests a common risk of environmental exposure to both bacteria, e.g. poor hygiene, rather than a causal relationship via reduced gastric acid production.

Physicians' prescribing practice for treatment of acute diarrhoea in young children in Jakarta.

Prescribing practices of physicians who treat acute childhood diarrhoea in Penjaringan, an urban district of Jakarta, were investigated utilising observations of clinical practice and interviews with physicians. One hundred thirty-four physicians registered in Penjaringan district in their practices; 122 physicians (91%) participated in the survey and 73 (55%) were included in the observational study. Reported and observed practices are compared. Results show that 78% of the physicians reported that they frequently prescribe Oralit, a solution for oral rehydration therapy (ORT). However, observation of clinical cases indicates that Oralit was prescribed less frequently than reported. Although most physicians believed viral infections were a common cause of diarrhoea, antibiotics were prescribed for children in 94 percent of observed cases. Antispasmodic drugs were also commonly prescribed. Factors associated with physicians' prescribing behaviour and practices regarding diarrhoeal diseases include type of practice, concepts about aetiology, perceptions about ORT and parental expectations. The observed discrepancy between knowledge and practice suggests the need for new ways to encourage physicians to prescribe Oralit and to limit use of antibiotics and antidiarrhoeals of doubtful efficacy.

Evaluation of a commercially available immunoglobulin M capture enzyme-linked immunosorbent assay kit for diagnosing acute dengue infections.

Recently, commercially available kits for the detection of anti-dengue virus (anti-DEN) immunoglobulin M (IgM) antibodies have been developed. These standardized assays have greatly enhanced our ability to effectively diagnose DEN infections. We conducted an evaluation of a test kit manufactured by MRL Diagnostics Inc. that is designed to detect anti-DEN IgM antibodies. Eighty paired samples from DEN-infected individuals were tested by the MRL DEN Fever Virus IgM Capture enzyme-linked immunosorbent assay (ELISA), the PanBio Duo ELISA, the PanBio Rapid Immunochromatographic Test (PRIT), and the IgM-IgG antibody capture (MAC/GAC) ELISA. All infections were confirmed by either PCR-assisted detection of DEN transcripts or by DEN isolation in C6/36 cells. Seventeen paired samples from individuals with no evidence of acute DEN infection were used as negative controls. The PRIT had the best sensitivity (100%), whereas the MAC/GAC ELISA and the PanBio Duo assay had the highest levels of specificity. The MRL ELISA and the PanBio Duo assay were the top performers when taking into consideration both sensitivity and specificity. All assays were able to detect DEN-specific antibodies in samples from patients with either primary or secondary infections, regardless of the infecting DEN serotype.

Congenital lactic acidosis: evaluation of the properties of the a199t natural variant of human pyruvate dehydrogenase e1alpha by in vitro mutation.

One cause of congenital lactic acidosis is a mutation in the E1 alpha-subunit of the pyruvate dehydrogenase multienzyme complex. Little is known about the consequences of these mutations at the enzymatic level. Here we study the A199T mutation by expressing the protein in Escherichia coli. The specific activity is 25% of normal and the K(m) for pyruvate is elevated by 10-fold. Inhibitors of lactate dehydrogenase might be a useful therapy for patients with such mutations.

Evaluation of a rapid assay for detection of Chlamydia trachomatis infections in outpatient clinics in South Kalimantan, Indonesia.

A multicenter cross-sectional survey was conducted comparing a commercially available chlamydial optical immunoassay (OIA) to the chlamydial ligase chain reaction (LCR). Endocervical samples from 415 outpatients visiting clinics from three hospitals in South Kalimantan, Indonesia, were evaluated. Relative to the LCR, the overall sensitivity and specificity of the OIA were 31.6 and 98.9%, respectively. The sensitivity of the OIA varied among the three hospital laboratories, ranging from 20 to 50%. The OIA performance was slightly lower on samples from patients attending dermatovenereology clinics than on samples from nondermatovenereology clinic patients. The results indicate that the OIA did not perform well compared to LCR.

Risk factors for transmission of foodborne illness in restaurants and street vendors in Jakarta, Indonesia.

In a previous risk factor study in Jakarta we identified purchasing street food as an independent risk factor for paratyphoid. Eating from restaurants, however, was not associated with disease. To explain these findings we compared 128 street food-vendors with 74 food handlers from restaurants in a cross-sectional study in the same study area. Poor hand-washing hygiene and direct hand contact with foods, male sex and low educational level were independent characteristics of street vendors in a logistic regression analysis. Faecal contamination of drinking water (in 65 % of samples), dishwater (in 91 %) and ice cubes (in 100 %) was frequent. Directly transmittable pathogens including S. typhi (n = 1) and non-typhoidal Salmonella spp. (n = 6) were isolated in faecal samples in 13 (7 %) vendors; the groups did not differ, however, in contamination rates of drinking water and Salmonella isolation rates in stools. Poor hygiene of street vendors compared to restaurant vendors, in combination with faecal carriage of enteric pathogens including S. typhi, may help explain the association found between purchasing street food and foodborne illness, in particular Salmonella infections. Public health interventions to reduce transmission of foodborne illness should focus on general hygienic measures in street food trade, i.e. hand washing with soap, adequate food-handling hygiene, and frequent renewal of dishwater.

Acute jaundice in Vientiane, Lao People's Democratic Republic.

Analysis of serum samples from patients with acute jaundice by means of enzyme-linked immunosorbent assay and polymerase chain reaction testing provided the first profile of this condition in Vientiane, Lao PDR, in 1995 and 1996. In a case-control, hospital-based study, evidence of acute infections due to hepatitis A and B viruses was found in 14% and 10% of cases, respectively. Hepatitis E virus, however, did not appear to contribute to clinically recognized acute jaundice. Similarly, antibody to hepatitis C virus was recognized in almost equal proportions of cases (8%) and controls (6%), thus representing probable background infections. The detection of hepatitis G virus marks the first report of this virus in Lao PDR. The large proportion (21%) of new leptospiral infections in cases without acute hepatitis A or B was notable. This finding suggests significant regional underreporting of leptospirosis as a cause of acute jaundice. The limited laboratory diagnostic capabilities for confirming a differential diagnosis of leptospirosis contribute to the lack of attention paid to this important health problem.

Clinical, molecular and serological outcomes of the Chikungunya outbreak in Grenada

OBJECTIVE: This study evaluated the clinical utility of Chikungunya (CHIKV) test results and clinical symptoms in patients with suspected CHIKV infection. DESIGN AND METHODS: Patients with CHIKV symptoms who presented at a health facility in Grenada during the recent outbreak had a CHIKV diagnostic test form completed by a health professional and a blood sample was drawn. The serum sample was stored at -80oC, shipped to the Naval Infectious Diseases Diagnostic Lab (NIDDL) on dry ice and tested for CHIKV and Dengue (DENV) using PCR real-time assay for viral RNA, and IgM detection by ELISA. RESULTS: Sera from more than 600 patients collected from mid September till mid October, 2014 were drawn and had a CHIKV diagnostic form completed. At the time of writing 112 patients sera have been tested at the NIDDL. 90% of patients had a positive test. PCR only was positive in 8% of patients. IgM only was positive in 83%, and both PCR and IgM were positive in 9% of patients. The major symptoms presented by patients were joint pain (84%), fever (81%), body pain (74%), headache (62%), chills (54%) and rash (49%). CONCLUSION: IgM testing detected 92% of test positive patients while PCR alone detected 17%. The IgM assay was clinically most useful. In an outbreak where dengue is ruled out and CHIKV is the cause, patients with the constellation of symptoms above could be considered positive for CHIKV infection with a 98% accuracy without confirmatory testing.

Establishment of laboratory testing capability for chikungunya virus in Grenada, West Indies

OBJECTIVE: Rapid diagnosis of Chikungunya (CHIKV) is important early in an epidemic. The study objective was to describe the process of implementing CHIKV testing capability in Grenada and to confirm the arrival of CHIKV on the main island of Grenada. DESIGN AND METHODS: In April, 2014 a collaborative study between the U.S. Naval Infectious Diseases Diagnostic Laboratory (NIDDL) and the clinical microbiology laboratory of St. George’s University (SGU) was started. SGU acquired essential instrumentation and patient samples, and NIDDL provided supplies and reagents, plus technical training experts. RESULTS: Personnel, supplies and equipment arrived in Grenada in August 2014. Set up of instruments and test validation were completed quickly. Initial CHIKV PCR and IgM tests found 3 PCR positive samples. The IgM assay found several presumptive positives that were unable to be confirmed due to ELISA instrument malfunction. PCR data indicated that CHIKV had arrived on the main island of Grenada no later than August, 2014. Based on arbovirus test demand, symptomatic patients began to increase in August, peaked in September, and tailed off during November. CONCLUSION: Both CHIKV tests were implemented and produced the first on-island reference test confirmation of CHIKV patients in Grenada. The most difficult part of this effort was training technologists in time to help with testing. Laboratory testing for CHIKV infection can be a challenge in developing states at a distance from support services. Collaborative links with established labs remains essential.