Immunochromatographic Method Research Articles

Comparison of analytical performance of the HbA1c assay on the fine care fluorescence immunochromatographic system.

Background: The Fluorescence Immunochromatographic method is a relatively new technique offering similar advantages to small clinical laboratories with the added value of reagents stability and increased precision. Here we aim to evaluate the Fluorescence Immunochromatographic method in terms of accuracy, precision, and linearity. Also, to compare its performance against the Bronate Affinity method. Methods: Reproducibility between the two methods was assessed by measuring ten prepared HbA1c reference materials with different concentrations. Additionally, the Bland-Altman Plot was also used to determine the comparability of measurements between the two methods within a 1.96 SD limit. Moreover, twenty repeated measurements of two HbA1c reference materials with different concentrations were carried out to assess accuracy and precision. Results: The “best fit values” after comparing ten measurements of the HbA1c reference materials between the two methods for the Slope of the linear regression model was 0.9764 ± 0.05566 with “Y” intercept of 0.5469 ± 0.4657, “X” intercept of -0.5601, and R2 of 0.9747. All ten results were inside the 1.96 limit of the Bland-Altman Plot. Similarly, comparison of two sets of HbA1c references materials, “Control N” and “Control P”, after twenty repeated analysis showed a Coefficient of Variation (CV) of 2.64% and 3.55% respectively. Conclusion: There is no significant difference between the two methods in detecting HbA1c. Method evaluation employing both a liner regression model of analysis as well as the Bland-Altman method showed no significant variance. This shows that the Fluorescent Immunochromatography instrument is suitable for detecting HbA1c in the clinically significant detection range.

Blood donation type and hepatitis B and C seroepidemiology at Hôpital Sominé DOLO de Mopti

Background: The increasing prevalence of hepatitis viral markers in developing countries results in a concern in the setting of blood transfusion safety. The aim of this study was to assess the prevalence of hepatitis viral markers among blood donors and their link with sociodemographic factors. Methods: We performed a retrospective study on blood donors screened for HBV and HCV. HBV and HCV serologies were performed by immunochromatographic methods using Alere DetermineTM HBs-Ag and SD BIOLOLINETM HCV-Ab. Data were analyzed by R 3.4.3. The logistic regression was fitted in univariate and multivariate analysis. Results: A total of 11.372 donors were enrolled. The overall prevalence was 10.4%. The specific prevalence of HBV and HCV were 8.9% and 1.5% respectively. The co-infection rate was 0.3%. HBV seropositivity proportion was significantly higher in family donors compared to volunteer (9,8% vs 6,0%, or = 1.71 [1.30-2.29] p < 0,001). HBV seropositivity was positively influenced by male sex aOR = 1,77 [1,4-2,26], p < 0.001 and unemployed aOR = 1.20 [1.03-1.40], p = 0.02 in multivariate analysis. Conclusion: Our data suggest that voluntary donors and particularly female which had an occupation had a lower prevalence of hepatitis viral markers.

Design and establish a rapid diagnostic kit for Viral Nervous Necrosis (VNN) disease using immunochromatography method

Background: Our investigation has approved that Viral Nervous Necrosis was the main causative agent for the outbreak in Mullet fish in the Caspian Sea. The disease was first reported in wild golden grey mullet (Liza auratus) in 2004. Since betanodaviruses are completely resistant to environmental conditions, it is possible to move easily via commercial activities. Also, the virus can introduce horizontally through contaminated water. So, the rapid diagnosis could be very useful in control, prevention and eradication programs. Also, in Monitoring & Surveillance, fast detection of the unknown outbreak could be vital and decrease their economic impacts. Methods and materials: For the preparation of antibody-gold particle conjugates, the concentration of Abs and pH of colloidal gold should be adjusted. A different volume of 1% sodium citrate was added to obtain the desired gold nanoparticles size in 30 nm. It was conjugated with an appropriate concentration Ab and in specific pH, and then this conjugation was blocked with serum albumin bovine. Afterwards, both various volumes of antibody-gold particle conjugates were coated on the conjugate pad and different concentration of pAb are used in a test line. To assure that the test strip will work correctly, the flow must also reach the control line, which has goat anti-mouse IgG (5 mg/ml) embedded in it. Both test and control lines were spotted on the membrane by using Helena applicator. Finally, the components of the strip were assembled and then the brain specimens of suspicious grey mullets and different dilution of the virus were evaluated. Results: With using of 1 μg/μl of pAbs on the test line and 4 μl per strip of gold- antibody conjugates on the conjugate pad, the designed strip was able to detect 103 TCID50/ml concentration of VNN. Then 50 brain samples that produced positive test results by PCR. The supernatant solution from brain homogenate of VNN-infected Liza aurata was confirmed negative for VNN by subsequent PCR an IHC testing, were evaluated using immunochromatography strip. It was recognized that the sensitivity and specificity of this kit are 64 and 100%, respectively. Conclusion: The accuracy of the immunochromatography kit was identical to both real-time PCR and IHC.

Seroprevalence of hepatitis b surface antigen infection among outpatients attending a tertiary care centre

Hepatitis B virus (HBV) infection is a chronic lifelong infection. Hepatitis B virus (HBV) infects more than 400 million people worldwide and is common cause of liver disease and liver cancer. In India, it is estimated that there are 40 million people chronically infected with HBV infection. The current study was conducted to estimate the burden of HBV infection in Bidar district of North Karnataka.The present study was conducted in the Department of Microbiology, Bidar Institute of Medical Sciences, Bidar. A Total 12,142 samples were tested by one step Immunochromatographic method for rapid detection of HBsAg to diagnose the infection.Among 12,142 samples tested, 284 samples were positive for HBsAg with prevalence rate of 2.34%. Age, gender and month wise distribution of infection was studied.This study highlights the prevalence and burden of HBV infection in Bidar district of North Karnataka. Which provide reference to future studies on the epidemiology of HBV infection in this region.

A simple and rapid immunochromatography test based on readily available filter paper modified with chitosan to screen for 13 sulfonamides in milk

In this study, we developed a novel, simple, rapid, and low-cost colloidal gold-based immunochromatography method, with filter paper replacing nitrocellulose membrane as the substrate. To obtain adequately immobilized protein, chitosan was used to functionalize the filter paper. After conditions and parameters were optimized, the novel immunochromatography method was applied for detection of sulfonamide residues in milk. Quantitative detection was accomplished using a smartphone and Photoshop software (Adobe Inc., San Jose, CA), allowing us to screen 13 sulfonamides with a limit of detection ranging from 0.42 to 8.64 μg/L and recovery ranging from 88.2 to 116.9% in milk. The proposed novel method performed similarly to the conventional method that uses a nitrocellulose membrane as the transport medium, and it had lower cost and better usability because of the inexpensive and easily available filter paper.

Frequency of dogs with anti Leishmania Chagasi antibodies by immunochromatographic assay (DPP) in the city of Sinop, Mato Grosso, Brazil

Dogs are considered the main reservoir for visceral leishmaniasis infection in humans, a serious disease that is a relevant public health problem due to its wide geographical distribution. To date, there haven’t been notification of Canine Visceral Leishmaniasis (CVL) in official health agencies of Sinop city, state of Mato Grosso. CVL diagnosis is difficult due to the variety of symptoms of the disease. In addition, animals may remain asymptomatic for life. In this study, the frequency of anti Leishmania chagasi antibodies was observed in 201 dogs referred to the clinical routine of the Veterinary Hospital of the Federal University of Mato Grosso, Campus of Sinop, during a 67 days period, through immunochromatographic method (DPP). Obtained results showed that seven animals were positive while one hundred ninety four were negative to the test. This result is relevant as far as it may increase the risk of CVL dispersion to non-endemic areas, highlighting the need of implementation effective measures to quickly detect Leishmania chagasi and prevent epidemiological situations.

Evaluation of an Antibody-Based Test for the Diagnosis of Chikungunya Infection

Background: The diagnosis of chikungunya is confusing due to similar clinical presentations of different viral illnesses.
 Objective: The purpose of the present study was to evaluate the accuracy of an antibody-based test for the diagnosis of chikungunya infection
 Methodology: This cross-sectional study was conducted at the Department of Internal Medicine, Bangabandhu Sheikh Mujib Medical University, Dhaka, from July to September 2017 when an outbreak of chikungunya occurred in Bangladesh. Chikungunya patients were evaluated by the IgM antibody test by immunochromatographic method (ICT) during both the early phase and the convalescent-phase. The sensitivity and specificity of the IgM antibody test were estimated considering the polymerase chain reaction (PCR) method as the gold standard.
 Result: The sensitivity and specificity of the IgM antibody test in the acute phase were 2.7 percent and 79.5 percent, respectively. In contrast, in the convalescence phase, sensitivity and specificity were 86.5 percent and 33.3 percent.
 Conclusion: Antibody-based testing was found not suitable for detecting chikungunya infection during the acute phase of the illness.
 Journal of Current and Advance Medical Research 2020;7(2): 68-72

GAMBARAN HASIL PEMERIKSAAN HEPATITIS B SURFACE ANTIGEN (HBsAg) PADA IBU HAMIL TRIMESTER SATU DI WILAYAH KERJA PUSKESMAS KOTA TENGAH

Hepatitis B is inflammation or infection in liver cells caused by viruses and can be acute or chronic. According to WHO 2018, hepatitis B is a quite serious problem. In addition, the incidence of hepatitis B infection often occurs in pregnant women and has a great potential of contracting the baby. HBsAg examination in first trimester pregnant women is an early detection of vertical transmission hepatitis B. Based on data from Gorontalo City Health Office, in 2017-2018 there was an increase in the number of pregnant women infected with the hepatitis B virus by 15 people. Purpose of this research was to determine the description of the results of HBsAg examination in first trimester pregnant women at the Community Healt Center of Central City.
 This research is a descriptive qualitative research using HBsAg examination and immunochromatography methods. The population in this research were 30 samples with a total sampling technique.
 The results of HBsAg examination showed that in the first trimester pregnant women a positive result was infected with hepatitis B virus is 1 person with a percentage of 3.3% and a negative result of 29 people or 96.7%. Based on the results of research it can be concluded that first trimester pregnant women with negative HBsAg status more than positive ones

Rapid detection of brucellosis using a quantum dot-based immunochromatographic test strip.

Novel diagnostic tools are a major challenge for brucellosis research, especially in developing countries. Herein, we established a handheld quantum dot (QD) immunochromatographic device for the fast detection of brucellosis antibodies in the field. Total bacterial protein extracted from Brucella 104M served as labelling and coating antigen. QD labelling and immunochromatography methods were used to optimise reaction conditions, labelling conditions, reaction temperature and storage temperature. QD test strips were employed to test brucellosis serum to determine their sensitivity, specificity and stability. Test strips were compared with Rose Bengal test, standard agglutination test and colloidal gold immunochromatographic assay. Labelled Brucella total protein displayed good specificity and no cross-reactivity. The concentration of labelled total bacterial protein was 3.9 mg/ml, the coating concentration was 2.0 mg/ ml, and the serum titre with the lowest detection sensitivity was 1:25. The optimal reaction temperature for the test strip was 25-30°C. The test strip was stable after storage at room temperature and the repeatability was high, with a coefficient of variation of 4.0%. After testing 199 serum samples, the sensitivity of the QD test strip was 98.53%, the specificity was 93.57%, and the coincidence rate with the standard agglutination test was 96.98%. The developed QD immunochromatographic method can be used for rapid detection and preliminary screening of brucellosis in the field.

Diagnosis of helicobacter pylori infection: problems and prospects

(HP) infection was performed using invasive and non-invasive methods. The study group consisted of 95 patients with dyspepsia. HP infection was detected in 47 patients (49.4 %). The expediency of using a set of diagnostic methods for detecting HP (PCR, immunochromatographic, bacteriological and method for determining urease activity) is proved. Most often (100 %) in patients HP infection was detected in biopsies using the PCR method. Somewhat less frequently it was detected when examining biopsies with an invasive biochemical method (AMA RUT Reader) (82 %) and fecal immunochromatographic method (83 %). Despite the fact that helicobacteriosis was detected bacteriologically in a small number of patients (24 %), this method is of particular value, since it allows you to assess the sensitivity to antimicrobial drugs and probiotics, and does not give false positive results.

Current Methods for Body Fluid Identification Related to Sexual Crime: Focusing on Saliva, Semen, and Vaginal Fluid.

Although, DNA typing plays a decisive role in the identification of persons from blood and body fluid stains in criminal investigations, clarifying the origin of extracted DNA has also been considered an essential task in proving a criminal act. This review introduces the importance of developing precise methods for body fluid identification. Body fluid identification has long relied on enzymatic methods as a presumptive assay and histological or serological methods as a confirmatory assay. However, because the latest DNA typing methods can rapidly obtain results from very small and even old, poorly preserved samples, the development of a novel corresponding body fluid identification method is required. In particular, an immunochromatographic method has been introduced to identify saliva and semen from sexual crimes. In addition, for vaginal fluid identification, attempts have been made in the past decade to introduce a method relying on body fluid-specific mRNA expression levels. At present, the development of molecular biological methods involving microRNA, DNA methylation, and resident bacterial DNA is ongoing. Therefore, in criminal investigations, body fluid identification is an essential task for correctly applying the results of DNA typing, although further research and development are required.

Detection and comparison of malaria by conventional, rapid immuno chromatographic and molecular technique

Background: Mosquito-borne diseases are the major concern in public health. Malaria is a protozoal disease caused by the parasites of the Genus Plasmodium - Plasmodium vivax, Plasmodium falciparum, Plasmodium malariae and Plasmodium ovale, which are transmitted by the bite of female Anopheles mosquitoes. Prevalence of malaria worldwide is about 35%. Malarial infections can be diagnosed by peripheral smear, rapid immunochromatography and molecular technique. This study is focused on Plasmodium species detection by using species specific primers for diagnosis. Aim and objective of this study was to detect and compare the Plasmodium species by conventional, rapid immunochromatographic and molecular technique in patients suspected for pyrexia of unknown origin (PUO) attending a tertiary care hospital.Methods: The study was carried out at SRM MCH and RC, Tamil Nadu, India, from January 2018 to February 2019 after the institutional ethical committee approval. This was an observational study. Malarial parasites were identified in peripheral blood smear by conventional method (thick smear and thin smear), rapid immunochromatographic technique (serological technique using whole blood) and molecular technique. DNA extraction from whole blood done by spine column technique. Amplification and gel documentation done.Results: Total 83 blood samples were collected from patients with clinical diagnosis of PUO. Out of 83 samples 5 (6%) samples were positive for Plasmodium vivax species by conventional method, rapid immunochromatographic and molecular technique.Conclusions: In this study, Plasmodium vivax is the species identified by conventional peripheral blood smear method, rapid immunochromatographic and molecular method.

Comparison of placental α microglobulin-1 protein assay (Amnisure) with speculum examination for the diagnosis of premature preterm rupture of membranes (PPROM): a clinical evaluation

In this study, we aimed to compare the clinical outcomes of Premature Preterm Rupture of Membranes (PPROM) cases diagnosed by classical speculum examination and by placental alpha microglobulin-1 protein (PAMG-1) assay. The medical records of all patients with singleton pregnancies that were diagnosed with PPROM were retrospectively reviewed. Singleton pregnancies with PPROM diagnosis that was confirmed either by direct visualisation of amniotic fluid leaking through the cervix or by placental alpha microglobulin-1 protein (PAMG-1) assay if no amniotic fluid leakage was documented were included in the study. Demographics, prenatal and postnatal characteristics were reviewed from the medical charts and were recorded. The study included 138 pregnancies with PPROM; 111 patients in clinical speculum examination group and 27 in PAMG-1 assay group. There were no significant differences in maternal and pregnancy characteristics between the clinical speculum examination and PAMG-1 assay groups. Foetal outcomes were comparable between clinical speculum examination and PAMG-1 assay groups. In the clinical speculum examination group, there were nine (8.1%) chorioamnionitis cases, however, there were no chorioamnionitis cases in the PAMG-1 assay group during the latency period (p = .21). Impact statement What is already known on this subject? Placental alpha microglobulin-1 protein assay uses immunochromatography method to detect trace amount of placental alpha microglobulin-1 protein in vaginal fluids and has high sensitivity and specificity for ROM diagnosis. However, to the best of our knowledge, the clinical outcome of ROM cases detected by classical speculum examination and by placental alpha microglobulin-1 protein assay has not been compared in the literature previously. What do the results of this study add? Although statistically insignificant, cases diagnosed by PAMG-1 assay had lower risk of chorioamnionitis during latency period. What are the implications of these findings for clinical practice and/or further research? Whether cases diagnosed by PAMG-1 assay represent a milder form of rupture of membranes than cases diagnosed by classical speculum examination group warrants further research.

Investigation of the rapid immunochromatographic test performance in the diagnosis of syphilis; comparison of four serological methods

AbstractObjectivesTo test the performance of the newly available rapid test for syphilis, we compared it withTreponema pallidumhemagglutination assay (TPHA). Additionally, we investigated the performance of rapid plasma reagin (RPR) and chemiluminescence microparticle immunoassays (CMIA) at our laboratory using TPHA as a gold standard.MethodsThe serum samples of 595 patients with the pre-diagnosis of syphilis were studied by four serological methods. The sensitivity, specificity, and predictive values of RPR, CMIA, and syphilis rapid test were assessed by utilizing TPHA as a gold standard for the diagnosis of syphilis.ResultsOf the patients, 6.2% (37/595) had positive RPR, 5.5% (33/595) had positive CMIA, 5.5% (33/595) had a positive rapid immunochromatographic method and 5% (30/595) had positive TPHA. When TPHA results were taken as the reference, the sensitivity of the rapid test for syphilis was 100%, the specificity was 99.5%, PPV was 90.9%, and NPV was 100.0%.ConclusionsIt was observed that the rapid test for syphilis used in the study was quite successful, its cost was appropriate, and the test was very fast and easy to apply. At the same time, the agreement between syphilis rapid test and TPHA was found to be excellent.

Rapid simultaneous detection of fumonisin B1 and deoxynivalenol in grain by immunochromatographic test strip

Mycotoxins, such as fumonisin B1 (FB1) and deoxynivalenol (DON), are toxic secondary metabolites produced by fungi. Herein, the simultaneous detection of FB1 and DON (both of which are water-soluble) in grain samples by immunochromatographic methods was used to probe whether the levels of these mycotoxins exceeded the limit of 1 mg/kg. Two types of immunochromatographic test strips (ICSs) were developed for this purpose, namely, those based on Au nanospheres (prepared via reduction with trisodium citrate) and Au nanoflowers (prepared by Au seed-mediated growth) as markers, with the respective sensitivities to both FB1 and DON equalling 20 and 5.0 ng/mL. The former ICS was used to detect FB1 and DON in grain (51 wheat samples and 18 maize samples), providing results consistent with those of high-performance liquid chromatography and enzyme-linked immunosorbent assays. The developed technique was suitable for the on-site screening of large-scale samples for FB1 and DON.

The Profiles of Hepatitis B and C Virus Infections in Prison-Assisted Citizens

Prison-assisted citizens or prisoners are one of the groups of people who are at high risk of Hepatitis B and C virus infections. The data on the prevalence of both viruses in prisoners in Indonesia are still limited. This study aims to determine the profiles of Hepatitis B and C virus infections in prison-assisted citizens in the Class IIA Narcotics Correctional Institution in Bandung Regency. A total of 30 prisoners were used as the samples in this study. All procedures performed in this study were following the applicable codes of ethics. The presence of surface antigens of Hepatitis B virus (HBsAg) in serums was detected using a qualitative sandwich Enzyme-Linked Immunosorbent Assay method. The existence of antibodies of the Hepatitis C virus (Anti-VHC) was detected using the immunochromatography method. The laboratory test results have shown five people (16.7%) were positive on the HBsAg test and one person (3.3%) was positive on the anti-VHC test. One case of Hepatitis B and C co-infection was also found in the prison-assisted citizens who were in the Class IIA Narcotics Correctional Institution in Bandung Regency. All the prisoners infected by the Hepatitis B and C viruses used injection drugs, had tattoos on their bodies and never received vaccinations.

Development of a Sensitive Immunochromatographic Method Using Lanthanide Fluorescent Microsphere for Rapid Serodiagnosis of COVID-19.

The SARS-CoV-2 infection that caused the COVID-19 pandemic quickly spread worldwide within two months. Rapid diagnosis of the disease and isolation of patients are effective ways to prevent and control the spread of COVID-19. Therefore, a sensitive immunofluorescent assay method was developed for rapid detection of special IgM and IgG of COVID-19 in human serum within 10 min. The recombinant nucleocapsid protein of 2019 novel coronavirus was used as capture antigen. Lanthanide, Eu(III) fluorescent microsphere, was used to qualitatively/semiquantitatively determine the solid phase immunochromatographic assay. A total of 28 clinical positive and 77 negative serum or plasma samples were included in the test. Based on the analysis of serum or plasma from COVID-19 patients and healthy people, the sensitivity and specificity of the immunochromatographic assay were calculated as 98.72% and 100% (IgG), and 98.68% and 93.10% (IgM), respectively. The results demonstrated that rapid immunoassay has high sensitivity and specificity and was useful for rapid serodiagnosis of COVID-19.

Screening for hepatitis b and c viral infections among pregnant women attending the Bolan Medical Complex Hospital and Sandeman Provincial Civil Hospital Quetta, Pakistan.

Infectious diseases caused by hepatitis B virus (HBV) and hepatitis C virus (HCV) are a worldwide burden to health, especially in developing countries. Pakistan has one of the highest levels of HBV and HCV infection, causing a severe health problem with significant challenges and priorities. To prevent vertical transmission of infection, regular screening of pregnant women for HBV and HCV is vital. Objectives: The objective of this study was to evaluate the prevalence of HBV and HCV virus in pregnant women having prenatal care. Study Design: Cross-Sectional study. Setting: Bolan Medical Complex Hospital and Sandeman Provincial Civil Hospital Quetta. Period: August 2017 to July 2018. Material & Methods: Blood serum samples were screened for HBV surface antigen (HBsAg) and for anti-HCV using immunochromatography methods. Results: A total of 12,209 pregnant women were tested over a period of one year (August 2017 to July 2018). The overall HBV infections frequency was 1.3% (95% CI 1.1-1.4%) and for HCV infections it was 0.6% (95% CI 0.6-0.7%). Whilst there was only small month-wise variation in the occurrence of HBV and HCV infections, HBV prevalence was highest in May (1.7%) and HCV prevalence was highest in August and December (0.8%). Conclusions: Screening of all pregnant women for HBV and HCV is essential for reducing and eliminating vertical transmission of infection. Risk factors for infection need to be avoided and managed properly.

Guidance for the management of adult patients with coronavirus disease 2019.

In December 2019, a novel coronavirus was identified in Wuhan City, Hubei Province, China and later the disease was named coronavirus disease 2019 (COVID-19). On March 11, 2020, the World Health Organization (WHO) officially announced that COVID-19 had reached global pandemic status. This article summarized the understanding of the etiology, pathogenesis, epidemiology, clinical characteristics, diagnosis, treatment, rehabilitation, and prevention and control measures of COVID-19 based on the available data and anti-epidemic experience in China.

Şanlıurfa’nın Halfeti ilçesindeki akut gastroenteritli çocuklarda rotavirüs ve adenovirüs sıklığı

Background/Aim: Enteric viruses (rotavirus, adenovirus, norovirus, etc.) have been reported as the most common cause of non-bacterial acute gastroenteritis. Rotavirus can cause severe fluid and electrolyte imbalance and the development of fatal complications during the course of the disease if not diagnosed and treated early. The aim of study was to evaluate the frequency, epidemiological characteristics, clinical and laboratory data of rotavirus and enteric adenovirus agents in children presenting with diarrhea. Material and Method: The records of patients with diarrhea between ages of 0-12 who applied to Halfeti State Hospital between July 2017 and July 2019 and diagnosed with acute gastroenteritis and whose rotavirus and enteric adenovirus tests (examined by immunochromatographic methods) were requested were evaluated retrospectively. The frequency of rotavirus and adenovirus gastroenteritis, demographic data, clinical status, laboratory findings of these patients; age groups, gender and seasonal distribution were examined. SPSS 25.0 package program was used for statistical analysis. Results: 640 patients were included in the study. The median age of the patients was 22 months and 57% were male. Rotavirus was detected in 40% of the patients and adenovirus was found in 10.2% of the patients. It was found that the hospitalization rates of children with positive rotavirus were higher than other groups. The children with rotavirus are mostly in winter; children with adenovirus mostly in spring; rotavirus and adenovirus negative children were admitted to the emergency department mostly during the summer months. Conclusion: Viral agents constitute an important part of acute gastroenteritis in our region. It was found that stool examinations were important in the diagnosis of rotavirus and adenovirus gastroenteritis causing high mortality and morbidity. We think that unnecessary antibiotic use will decrease with appropriate treatment and the incidence of the disease and hospitalization cost burden will decrease with increasing rotavirus vaccination rates for Turkey.