Dengue virus detection in Lao PDR and Colombia: Comparative evaluation of PCR tests.

OBJECTIVES: Dengue virus (DENV) detection by polymerase chain reaction (PCR) facilitates diagnosis of dengue fever, which is the most frequent arboviral disease globally. Two studies were performed in countries with high dengue incidence, to assess the diagnostic performance of different PCR techniques. METHODS/RESULTS: Two hundred and seventy-nine acute phase blood samples from febrile patients were analyzed for DENV by the RealStar Dengue RT-PCR kit (Altona Diagnostics) as gold standard in comparison with the Tropical Fever Core multiplex PCR (Fast Track Diagnostics). In total, 102 samples collected in Savannakhet Province (Lao PDR, Southeast Asia) in 2013 and 35 samples from Valledupar (Colombia, South America) tested positive for DENV by RealStar RT-PCR. In comparison, the Tropical Fever Core multiplex PCR detected 65.0% (65/102) and 68.6% (24/35) of these samples as positive for DENV in Savannakhet and Valledupar, respectively. Diagnostic sensitivity of the multiplex PCR strongly correlated with viral load. A subset of DENV PCR-confirmed samples was additionally tested by BNITM in house Dengue Type RT-PCR in comparison with two commercial test kits (RealStar Dengue Type RT-PCR [Altona Diagnostics], Dengue differentiation PCR [Fast Track Diagnostics]). The leading dengue serotype in Savannakhet was DENV-3 (58% [29/50]), while DENV-1 (53.8% [14/26]) was the predominant serotype found in samples collected in Valledupar by BNITM-type PCR. However, three DENV serotypes were circulating in Valledupar and in Savannakhet. In 2015, additional studies found predominantly DENV-4 (71% [12/17]) in Savannakhet. CONCLUSIONS: Both studies emphasized that routine diagnostics in both regions will benefit from an expanded use of highly sensitive pan-dengue PCRs.

Assessment of diagnostic and analytic performance of the SD Bioline Dengue Duo test for dengue virus (DENV) infections in an endemic area (Savannakhet province, Lao People's Democratic Republic).

BACKGROUND: Rapid tests detecting both dengue virus (DENV) NS1 antigen and anti-DENV IgM and IgG antibodies facilitate diagnosis of dengue fever (DF) in resource-poor settings. METHODOLOGY/PRINCIPAL FINDINGS: 92 acute phase serum samples from patients with a PCR-confirmed DENV infection collected in Lao People's Democratic Republic (Lao PDR) in 2013 and 2015 were analyzed with the SD Bioline Dengue Duo test. A subset of 74 samples was additionally tested with the Platelia NS1 antigen test, the Panbio DENV µ-capture ELISA and the Panbio DENV IgG ELISA. IgM seroconversion was assayed using follow-up samples of 35 patients collected in the convalescent phase. 57.6%, 22.8% and 44.6% of acute phase serum samples tested positive in the SD Bioline Dengue Duo NS1, IgM, and IgG test, respectively. Diagnostic sensitivity of the SD Bioline Dengue Duo NS1 test strongly correlated with viral load, decreased rapidly over the acute phase of the disease, and was significantly reduced in presence of high anti-DENV IgG antibody titers resulting from secondary DENV infection. While a good concordance (Cohen's kappa 0.78) was found between the SD Bioline Dengue Duo NS1 test and the Platelia NS1 antigen ELISA, both the SD Bioline Dengue Duo IgM and IgG test displayed a significantly lower sensitivity than the corresponding ELISA tests. CONCLUSIONS/SIGNIFICANCE: The SD Bioline Dengue Duo test is a valuable tool for diagnosis of DENV infections especially when analyzing early acute phase samples with high viral load. Nevertheless, in endemic areas, where secondary flavivirus infections are common, diagnostic sensitivity of the NS1 and IgM test components may be compromised.

Morbidity assessment of Opisthorchis viverrini infection in rural Laos: I. Parasitological, clinical, ultrasonographical and biochemical findings.

BACKGROUND: Infections with the food-borne trematode Opisthorchis viverinni are common in Southeast Asia. In Lao PDR alone, two million people are supposed to be infected. Opisthorchiasis may cause severe liver disease, eventually leading to cholangiocarcinoma. The objective of this study is to assess the eating habits, complaints, symptoms, signs and ultrasonographical findings in three different areas of Savannakhet Province. METHODS: Study participants were recruited in Lahanam village in the flood-prone lowland of, Sonkhone district, Savannakhet Province (group A); in Non Somboon village, a community located on a hilly plateau in the same district (group B); and in staff of Savannakhet Province Hospital, Savannakhet town (group C). Eating habits, complaints and symptoms were recorded by standardized structured questionnaires. Participants were thoroughly examined clinically, and ultrasonography was performed. O. viverrini eggs were looked for in stool and in duodenal fluid. An array of biochemical and haematological parameters potentially related to liver disease was determined. Group A consisted of 45, group B of 31 and group C of 18 individuals. RESULTS: Eating habits were similar in the three groups, except that participants from group C tended to consume less high-risk types of fish dishes and more frequently ate beef and pork. Average intensity of infection (eggs per gram of stool) was low, but significantly higher in group A than in group B and C (p < 0.001). Medical history and complaints were similar in the three groups. Ultrasonography did not reveal any bile duct pathology. The only pathological finding was a slight elevation of ASAT and gamma-GT in a few participants in groups A and B. CONCLUSIONS: The study shows that eating habits favouring the infection with O. viverrini are common in south Laos. Although the average intensity of infection was low, there was a significant difference between the groups, paralleling slightly different eating habits. Clinically, this corresponded to a paucity of liver disease-associated complaints and signs. The low intensity of infection probably explains why no alterations of bile ducts were detectable by ultrasonography.