Diagnosis Of Cholera Research Articles

Seroepidemiologic Survey of Epidemic Cholera in Haiti to Assess Spectrum of Illness and Risk Factors for Severe Disease

To assess the spectrum of illness from toxigenic Vibrio cholerae O1 and risk factors for severe cholera in Haiti, we conducted a cross-sectional survey in a rural commune with more than 21,000 residents. During March 22–April 6, 2011, we interviewed 2,622 residents ≥ 2 years of age and tested serum specimens from 2,527 (96%) participants for vibriocidal and antibodies against cholera toxin; 18% of participants reported a cholera diagnosis, 39% had vibriocidal titers ≥ 320, and 64% had vibriocidal titers ≥ 80, suggesting widespread infection. Among seropositive participants (vibriocidal titers ≥ 320), 74.5% reported no diarrhea and 9.0% had severe cholera (reported receiving intravenous fluids and overnight hospitalization). This high burden of severe cholera is likely explained by the lack of pre-existing immunity in this population, although the virulence of the atypical El Tor strain causing the epidemic and other factors might also play a role.

Cholera with severe renal failure in an Italian tourist returning from Cuba, July 2013

In July 2013, an Italian tourist returning from Cuba was hospitalised in Trieste, Italy, for cholera caused by Vibrio cholerae O1 serotype Ogawa with severe renal failure. An outbreak of cholera was reported in Cuba in January 2013. Physicians should consider the diagnosis of cholera in travellers returning from Cuba presenting with acute watery diarrhoea.

Evaluation of a Rapid Test for the Diagnosis of Cholera in the Absence of a Gold Standard

BackgroundEarly detection and confirmation of cholera outbreaks are crucial for rapid implementation of control measures. Because cholera frequently affects regions with limited laboratory resources, rapid diagnostic tests (RDT) designed for field conditions are important to enhance rapid response. Stool culture remains the “gold standard” for cholera diagnosis; however, its lack of sensitivity may lead to underestimation of test specificity. We evaluated the Crystal VC® immunochromatographic test (Span Diagnostics, India) for cholera diagnosis using a modified reference standard that combines culture-dependent and independent assays, or a Bayesian latent class model (LCM) analysis.Methodology/Principal FindingsThe study was conducted during a cholera epidemic in 2008, in Lubumbashi, Democratic Republic of Congo. Stools collected from 296 patients were used to perform the RDT on site and sent to Institut Pasteur, Paris, for bacterial culture. In comparison with culture as the gold standard, the RDT showed good sensitivity (92.2%; 95% CI: 86.8%–95.9%) but poor specificity when used by a trained laboratory technician (70.6%; 95% CI: 60.7%–79.2%) or by clinicians with no specific test training (60.4%, 95% CI: 50.2%–70.0%). The specificity of the test performed by the laboratory technician increased to 88.6% (95% CI: 78.7–94.9) when PCR was combined with culture results as the reference standard, and to 85.0% (95% CI: 70.4–99.2), when the Bayesian LCM analysis was used for performance evaluation. In both cases, the sensitivity remained high.ConclusionUsing an improved reference standard or appropriate statistical methods for diagnostic test evaluations in the absence of a gold standard, we report better performance of the Crystal VC® RDT than previously published. Our results confirm that this test can be used for early outbreak detection or epidemiological surveillance, key components of efficient global cholera control. Our analysis also highlights the importance of improving evaluations of RDT when no reliable gold standard is available.

Chemoprophylaxis in contacts of patients with cholera: systematic review and meta-analysis.

IntroductionThere is a pressing need for effective measures to prevent the spread of cholera. Our systematic review assesses the effects of chemoprophylaxis in preventing cholera among exposed contacts.Methods and FindingsWe considered published and unpublished reports of studies up to July 2011. For this we searched: PubMed (1966 to July, 2011), Embase (1980 to July 2011), Cochrane Central Register of Controlled Trials (6; 2011), LILACS (1982 to July, 2011), the International Clinical Trials Registry Platform (July 2011) and references of identified publications. We included controlled clinical trials (randomized and non-randomized) in which chemoprophylaxis was used to prevent cholera among patient contacts. The main outcome measures were hospitalization and laboratory diagnosis of cholera in contacts for cholera patients. We assessed the risk of bias. We identified 2638 references and these included 2 randomized trials and 5 controlled trials that added up to a total of 4,154 participants. The risk of bias scored high for most trials. The combined results from two trials found that chemoprophylaxis reduced hospitalization of contacts during the follow-up period by 8–12 days (2826 participants; RR 0.54 95% CI 0.40–0.74;I2 0%). A meta-analysis of five trials found a significant reduction in disease among contacts with at least one positive sample who received chemoprophylaxis during the overall follow-up (range 4–15 days) (1,414 participants; RR 0.35 95% CI 0.18–0.66;I2 74%). A significant reduction in the number of positive samples was also found with chemoprophylaxis (3 CCT; 6,918 samples; RR 0.39 95% CI 0.29–0.51;I2 0%).ConclusionOur findings suggest that chemoprophylaxis has a protective effect among household contacts of people with cholera but the results are based on studies with a high risk of bias. Hence, there is a need for adequate reliable research that allows balancing benefits and harms by evaluating the effects of chemoprophylaxis.

Preparing Health Care Workers for a Cholera Epidemic, Dominican Republic, 2010

Preparing Health Care Workers for a Cholera Epidemic, Dominican Republic, 2010

Use of Filter Paper as a Transport Medium for Laboratory Diagnosis of Cholera under Field Conditions

Confirmation of a cholera epidemic is based on bacteriological identification of the agent and requires the sending of samples to a culture laboratory, often in countries with limited resources. Comparison of the use of filter paper with the use of Cary-Blair reference medium for stool transport showed that this simple transport medium is appropriate for the recovery of Vibrio cholerae.

Dry-reagent gold nanoparticle-based lateral flow biosensor for the simultaneous detection of Vibrio cholerae serogroups O1 and O139

Cholera is a communicable disease caused by consumption of contaminated food and water. This potentially fatal intestinal infection is characterised by profuse secretion of rice watery stool that can rapidly lead to severe dehydration and shock, thus requiring treatment to be given immediately. Epidemic and pandemic cholera are exclusively associated with Vibrio cholerae serogroups O1 and O139. In light of the need for rapid diagnosis of cholera and to prevent spread of outbreaks, we have developed and evaluated a direct one-step lateral flow biosensor for the simultaneous detection of both V. cholerae O1 and O139 serogroups using alkaline peptone water culture. Serogroup specific monoclonal antibodies raised against lipopolysaccharides (LPS) were used to functionalize the colloidal gold nanoparticles for dual detection in the biosensor. The assay is based on immunochromatographic principle where antigen–antibody reaction would result in the accumulation of gold nanoparticles and thus, the appearance of a red line on the strip. The dry-reagent dipstick format of the biosensor ensure user-friendly application, rapid result that can be read with the naked eyes and cold-chain free storage that is well-suited to be performed at resource-limited settings.

Two Different Diagnosis Methods For The detection of cholera toxin production From Vibrio cholerae isolated from different areas in Iraq

Fifty isolates of Vibrio cholerae obtained from different areas in Iraq from patients with acute secretory diarrhea were diagnosed serologically in central public health laboratory(CPHL), Two different methods were used for detection cholera toxin production ,the first based on the degradation of Nicotin Amide Adinine Dinucleotide( NAD) by Cholera toxin (CT). and the second was by detection CT gene by polymerase chain reaction (PCR).All Vibrio cholerae were cultured in alkaline peptone water, out of 50 isolates 46 (92%) were positive when we used NAD for the detection of cholera toxin(CT).Whereas all isolates gave positive results by PCR. From these data we conclude that the degradation of NAD by CT is simple and can be carried out in small laboratories, it is also easy to perform, and gives reproducible results, while PCR provides a more sensitive and specific assay for rapid diagnosis of cholera than currently available methods.

Evaluation of a Rapid Immunochromatographic Dipstick Kit for Diagnosis of Cholera Emphasizes Its Outbreak Utility

We evaluated the Crystal VC, a commercially produced dipstick, for the rapid detection of Vibrio cholerae serotypes O1 and O139 directly from the stool samples of hospitalized diarrheal patients using the conventional bacteriological method as gold standard. The sensitivity and specificity of the dipsticks were about 92 and 73%, respectively. Introduction of the PCR-based method along with the classical bacteriological method as the gold standard for the evaluation of a kit may improve the sensitivity and specificity of the assay. The dipstick method requires minimal technical skill, and the test can be read in about 10 min. This dipstick test has the potential to act as an early warning system for cholera in many developing countries, especially during the start of an outbreak, which would ultimately lead to a decrease in the spread of the disease as well as the case fatality rate. Furthermore, the use of a rapid detection test will improve surveillance and thus reduce the burden of disease estimates, especially in remote settings.

Field evaluation of Crystal VC® Rapid Dipstick test for cholera during a cholera outbreak in Guinea‐Bissau

To evaluate performance characteristics and ease of use of the new commercially available Crystal VC Rapid Dipstick (VC) test (Span Diagnostics, India) for Vibrio cholerae O1 and O139. Whole stool was collected from patients presenting to a hospital cholera ward during a 2008 epidemic in Guinea-Bissau. The VC test on stool samples was conducted on-site; samples were subsequently stored in Cary-Blair transport media and sent to the Centers for Disease Control and Prevention for diagnostic testing by culture and polymerase chain reaction (PCR). In addition, four local laboratory technicians who were unfamiliar with the test were provided with stool samples, the VC test kit, and simple written instructions and asked to perform the test and interpret results. A total of 101 stool specimens were collected and tested. Compared with PCR, the test was 97% sensitive and 71-76% specific. Laboratory technicians in Bissau performed the test and interpreted results correctly using only simple written instructions. The VC test may be useful for cholera diagnosis in outbreak situations where laboratory capacity is limited.

Index of Suspicion

Index of Suspicion

The impact of socio-economic status and sanitation levels on the prevalence of diarrhoeal diseases in the Akim Oda area of Ghana

A survey was conducted between August and October, 1998 in Akim Oda area of Ghana to relate socio-economic differences, as measured by income, education, occupation and housing, among residents in three communities to diarrhoea prevalence. There was clinical and laboratory diagnosis of cholera and administration of structured interview-administered questionnaires to find from residents their demographic, socio-economic, health practices and status, and to seek their opinions on cholera transmission, control and prevention. The survey revealed that residents from Old-town and Aduasa belong to low social class and was reflected in their high illiteracy, unemployment rates and associated poor sanitation. As such, they had the highest prevalence of diarrhoeal diseases. Contrarily, residents from Quarters residential area were found to belong to high social class, which was reflected in their high literacy, employment rates and good sanitation. As such, they had no case of diarrhoea. The underlying factors responsible for this epidemic are discussed. Name of institutions where the work was done: Akim Oda government hospital and Korle-Bu Public Health and Reference laboratory.

Choléra

Cholera is an acute intestinal infection that has reached pandemic proportions and presents a major international health concern. Every year, more than 100000 cholera cases and 2000-3000 deaths are officially reported to WHO. The real figures for cholera are thought to be much higher, however, due to underreporting and other limitations of surveillance systems. Cholera is caused by two serogroups (O1 and O139) of a gram-negative bacterium, Vibrio cholerae. Cholera toxins cause a massive outpouring of electrolyte-rich isotonic fluid into the bowel and can lead to volume depletion and shock. In poor sanitary and individual hygiene conditions, the massive release of cholera vibrios into the environment intensifies and exacerbates cholera epidemics, which thus serve as clear markers of poverty and lack of basic sanitation. Rehydration therapy, either intravenous or oral, considerably decreases the number of deaths. The WHO recommends antibiotics for cholera cases with severe dehydration. If left untreated, cholera has a 25-50% mortality rate. Treatment reduces this to less than 1%. Bacteriological diagnosis of cholera is reasonably easy because cholera bacteria are abundant in stool. Epidemics, however, often occur in areas with either limited or no laboratory facilities. A rapid and accurate diagnosis of cholera is essential to mobilize resources for treatment and containment of the epidemic. Therefore, the Pasteur Institute has developed a rapid diagnostic test based on a one-step immunochromatographic technique, which should be commercially available soon. To date, two oral cholera vaccines have been shown to offer good (more than 70%) short-term (one year) protection. WHO recommends these vaccines as an additional public health tool to be implemented with the standard cholera control measures, including provision of safe drinking water and adequate sanitation. Nonetheless, a cholera vaccine that can offer a long-term protection for all age groups, including children younger than five years, is still needed.

Field evaluation of a rapid immunochromatographic dipstick test for the diagnosis of cholera in a high-risk population

BackgroundEarly detection of cholera outbreaks is crucial for the implementation of the most appropriate control strategies.MethodsThe performance of an immunochromatographic dipstick test (Institute Pasteur, Paris, France) specific for Vibrio cholerae O1 was evaluated in a prospective study in Beira, Mozambique, during the 2004 cholera season (January-May). Fecal specimens were collected from 391 patients with acute watery nonbloody diarrhea and tested by dipstick and conventional culture.ResultsThe overall sensitivity and specificity of the rapid test compared to culture were 95% (95% confidence interval [CI]: 91%–99%) and 89% (95% CI: 86%–93%), respectively. After stratification by type of sample (rectal swab/bulk stool) and severity of diarrhea, the sensitivity ranged between 85% and 98% and specificity between 77% and 97%.ConclusionThis one-step dipstick test performed well in the diagnosis of V. cholerae O1 in a setting with seasonal outbreaks where rapid tests are most urgently needed.

Evaluation of three rapid diagnostic tests for cholera: does the skill level of the technician matter?

To evaluate SMART, Medicos Dip Stick and an Institut Pasteur (IP) cholera dipstick tests for accuracy and ease of use. Every 50th patient presenting with diarrhoea at ICDDR,B between 1 April 2003 and 30 November 2003 was enrolled. The rapid diagnostic tests were performed by field and laboratory technicians, and sensitivity (Se), specificity (Sp), positive (PPV) and negative (NPV) predictive values calculated. We isolated Vibrio cholerae O1 from 116 (38%) of 304 patients. The Se, Sp, PPV and NPV of the SMART test were 58%, 95%, 84% and 84% for field technicians, and 83%, 88%, 83% and 88% for laboratory technicians. The Se, Sp, PPV and NPV of the IP dipstick test were 93%, 67%, 63% and 94% for field technicians, and 94%, 76%, 70% and 95% for laboratory technicians. The Se, Sp, PPV and NPV of the Medicos test were 84%, 79%, 71% and 90% for field technicians, and 88%, 80%, 72% and 92% for laboratory technicians. A high proportion of indeterminates (30%) hampered the performance of the SMART test. The IP dipstick had the highest Se, irrespective of technician skill level. The IP dipstick is the most appropriate rapid diagnostic assay for the detection of V. cholerae O1 in locations where the skill level of personnel may be low, such as remote areas or refugee camp settings. High cost may limit the utility of any diagnostic test in the developing world.

Desempeño de los sistemas Cholera-SMART® y Pathogen-Detection-Kit® en el diagnóstico rápido del cólera

To compare the performance of two rapid systems for the diagnosis of cholera with the culture method, and to propose a strategy for improving the specificity and sensitivity of these systems and reducing the costs involved in making a diagnosis. The following institutions participated in the study: the National Bacteriology Referral Center (Centro Nacional de Referencia en Bacteriologia, CNRB) of the Costa Rican Institute for Research and Teaching in Nutrition and Health (Instituto Costarricense de Investigacion y Ensenanza en Nutricion y Salud, INCIENSA) and various hospitals in the provinces of Alajuela, Guanacaste and San Jose, in Costa Rica. A total of 237 feces samples were used to asses the performance of two tests for the rapid detection of Vibrio cholerae 01: the Pathogen Detection Kit (PDK, Intelligent Monitoring Systems, Gainesville, Florida, USA) and Cholera-SMART (New Horizons Diagnostics Corp., Columbia, Maryland, USA), both when applied directly (direct SMART and direct PDK) and when applied to specimens cultured in broth-enriched medium for 6 hours (SMART-6 and CPK-6) and for 18 hours (SMART-18 and PDK-18) at 37 degrees C in alkaline peptone water. Liquid and partially formed stools were cultured and examined by means of the rapid direct test; when the initial result was negative, the tests were repeated after culture for periods of 6 and 18 hours. Rectal and fecal swabs were obtained from feces cultured in enriched-broth medium for 6 and 18 hours. In addition, we studied the sensitivity of the rapid testing systems by using pure cultures of V. cholerae 01 (strain SOS-833, CNRB, Costa Rica) that were incubated for 18 to 24 hours, and we assessed the usefulness of observing motility under the microscope in order to rationalize the use of rapid methods. The sensitivity of the direct SMART test and of the direct PDK test was 100% when samples obtained from liquid and partially formed stools and from the intestinal contents of dead bodies were used. With these samples, the direct SMART procedure showed a specificity of 100%, whereas the direct PDK procedure showed a specificity that ranged from 85.7% to 77.4%, depending on the type of sample. False positives obtained with the direct PDK method turned out to be negative with PDK-6 and PDK-18. Among the rectal and fecal swabs of persons with and without diarrhea or who had received prior treatment with antibiotics, three results that were negative with the SMART-6 procedure and two that were negative with the PDK-6 procedure turned out to be positive with the SMART-18 and PDK-18 procedures, respectively. Both systems showed excellent concordance (kappa index above 0.9) throughout. Both systems were sensitive to 6 x 10(7) colony-forming units per milliliter (cfu/mL), which was concordant with the microscopic observation of 10 microorganisms or more per field with the type of motility that characterizes vibrios (at 1000 x magnification). Samples having fewer than 10 microorganisms with the motility that characterizes vibrios had concentrations between 6 x 10(3) and 6 x 10(6) cfu/mL and became positive only after incubation in enriched-broth medium for 6 to 18 hours. We propose a strategy for diagnosing the presence of V. cholerae 01 infection in less time than it takes with traditional methods, with positive and negative predictive values of 100%. The SMART and PDK systems make it possible to accurately diagnose cholera quickly, don't require sophisticated equipment or highly qualified technical personnel, and perform satisfactorily in field conditions. Through the proposed strategy, it becomes possible to improve the specificity and sensitivity of these systems and to reduce the cost of making a diagnosis, thus making them suitable for use in cholera surveillance in low-income settings where this disease is a serious public health problem.

Use of dipsticks for rapid diagnosis of cholera caused by Vibrio cholerae O1 and O139 from rectal swabs.

We evaluated the recently developed dipsticks for the rapid detection of Vibrio cholerae serotypes O1 and O139 from rectal swabs of hospitalized diarrheal patients after enrichment for 4 h in alkaline peptone water. The sensitivity and specificity of the dipsticks were above 92 and 91%, respectively. The dipsticks represent the first rapid test which has been successfully used to diagnose cholera from rectal swabs, and this would immensely improve surveillance for cholera, especially in remote settings.

Validation of clinical and epidemiological diagnosis criterion for confirming cholera

To validate the clinical and epidemiological diagnosis criterion for confirming cholera suspect cases. The study comprised 2,687 and 716 patients admitted with diarrhea to a public hospital in Macei , Brazil, in 1992 and 1997 respectively. Culture of V. cholerae O1 (Koch, 1884) in TCBS-agar was performed and the clinical and epidemiological criterion was applied and compared to the gold standard. The statistical analysis was carried out in two age-groups: less than five years old and five years old or more. There were studied 833 patients, of which 517 in 1992 and 316 in 1997; 72 aged less than five years and 761 aged five years or more. In those aged less than five years in 1992, sensitivity was 40% and specificity 84.6%. For this same age-group in 1997, sensitivity was 28.6% and specificity 62.5%. In patients aged five years or more in 1992, sensitivity was 99% and specificity 1.2%. For this same age-group in 1997, sensitivity was 86.9% and specificity 8.7%. The higher sensitivity of the clinical and epidemiological cholera diagnosis criterion in patients aged five years or more found in the study support its application during epidemics periods. In periods of lower incidence, all cases need to be confirmed using laboratory methods.

New insights on the emergence of cholera in Latin America during 1991: the Peruvian experience.

After a century of absence, in late January 1991, Vibrio cholerae invaded the Western Hemisphere by way of Peru. Although a number of theories have been proposed, it is still not understood how that invasion took place. We reviewed the clinical records of persons attending hospital emergency departments in the major coastal cities of Peru from September through January of 1989/1990 and 1990/1991. We identified seven adults suffering from severe, watery diarrhea compatible with a clinical diagnosis of cholera during the four months preceding the cholera outbreak, but none during the previous year. The patients were scattered among five coastal cities along a 1,000 km coastline. We postulate that cholera vibrios, autochthonous to the aquatic environment, were present in multiple coastal locations, and resulted from environmental conditions that existed during an El Nino phenomenon. Once introduced into the coastal communities in concentrations large enough for human infection to occur, cholera spread by the well-known means of contaminated water and food.

Rapid diagnosis of cholera caused by Vibrio cholerae O139.

Hybridomas secreting specific monoclonal antibodies (MAbs) to Vibrio cholerae serogroup O139 were produced. Six monoclones (hybridomas) secreting MAbs specific only to lipopolysaccharide of V. cholerae O139 strains and which did not cross-react to 137 strains of other enteric microorganisms were obtained. These clones were designated 12F5-G11, 12F5-G2, 15F5-H5, 5B9-F8, 14C9-D2, and 6D2-D8. The immunoglobulin (Ig) heavy chain isotypes secreted by these clones were IgG2b, IgG2b, IgG2b, IgM, IgG2b, and IgG3, respectively. Clone 12F5-G11 was selected for mass production of MAb, which was used as a detection reagent in the antigen detection assay for diagnosis of cholera caused by V. cholerae O139, and this assay was compared to the conventional bacterial isolation method. Five batches of rectal swab cultures in alkaline-peptone water were collected from 6,497 patients with watery diarrhea. These were 6,310 patients admitted to Bamrasnaradura Infectious Diseases Hospital, 16 patients from Krung Thon Hospital, 78 patients from Bangkok Children's Hospital, 19 patients from Karen refugee camps, and 74 Indian patients from the National Institute of Cholera and Enteric Diseases, Calcutta, India. The V. cholerae O139 isolations from the rectal swab cultures and the antigen detection assays (i.e., the MAb-based dot-blot ELISA) were performed by different persons of different laboratories, and the results were revealed after all specimens had been tested. Of the 6,497 samples tested, the dot-blot ELISA correctly identified 42 of 42 V. cholerae O139-positive samples and gave a result of positive for three samples which were culture negative for V. cholerae O139. The diagnostic sensitivity, specificity, and efficacy of the dot-blot ELISA were 100, 99.95, and 99.26%, respectively. The ELISA is easy to perform and relatively inexpensive. It can test multiple samples at a single time, does not require special equipment, and does not produce great quantities of contaminated waste. Most of all, it reduces the diagnostic time from at least 2 days for the bacterial isolation to less than 90 min. The assay is recommended as a rapid screening test of cholera cases caused by V. cholerae O139.