A randomized controlled trial to evaluate the effect of influenza vaccination and probiotic supplementation on immune response and incidence of influenza-like illness in an elderly population in Indonesia.

AIM: To investigate the effect of influenza vaccination with or without probiotic supplementation on the immune response and incidence of influenza-like illness (ILI) in the elderly. METHODS: A randomized double-blind, placebo-controlled trial with a modified factorial design was conducted in 554 healthy elderly subjects aged 67 ± 5.6 (ranging from 60-90) years old in the Primary Health Care Center (Puskesmas area) of the Pulo Gadung District East Jakarta. Subjects received either a trivalent influenza vaccine or placebo at the start of the study, and a probiotic supplement (Lactobacillus helveticus R0052 and Lactobacillus rhamnosus R0011) or a placebo for 6 months. Subjects were randomly assigned into four intervention groups: influenza vaccine and probiotics (n = 141), influenza vaccine and placebo (n = 136), placebo and probiotics (n = 140), and both placebo (n = 137). The primary outcome was ILI incidence within 6 months. The secondary outcomes were seroprotection and seroconversion rates at 1, 4, and 6 months after administering the interventions. RESULTS: This study showed that the trivalent influenza vaccine increased seroprotection (RR 3.6 [95%CI 2.92-4.47]; p<0.010) and seroconversion (RR 29.8 [95%CI 11.1-79.5]; p<0.010) rates 1 month after vaccination in elderly people while the probiotic supplement did not alter influenza antibody titers (p = 1.000 and p = 0.210). The relative ILI incidence risk was similar between vaccinated and non-vaccinated groups, as well as in the probiotic group compared to the non-probiotic group. CONCLUSION: The tested trivalent influenza vaccine significantly induced seroprotection and seroconversion in the vaccinated subjects, while probiotics administration did not influence these parameters. Vaccinated individuals displayed a similarly low ILI incidence as those in the Control Group. However, the observed trend towards a reduction of ILI incidence with probiotics supplementation warrants further assessments in a larger, at-risk population. CLINICAL TRIAL REGISTRY NUMBER: NCT03695432.

Feasibility of Qualitative Testing of BCR-ABL and JAK2 V617F in Suspected Myeloproliperative Neoplasm (MPN) Using RT-PCR Reversed Dot Blot Hybridization (RT-PCR RDB).

BACKGROUND: Defining the presence of BCR-ABL transcript in suspected myeloproliferative neoplasm is essential in establishing chronic myeloid leukemia. In the absence of BCR-ABL, the conventional diagnostic algorithm recommends JAK2 V617F mutation testing to support diagnosis of other MPN diseases such as polycythemia vera, essential thrombocythemia, and primary myelofibrosis. In certain cases of thrombocythemia, simultaneous upfront testing of both BCR-ABL and JAK2 may be desirable. We wanted to test the feasibility of multiplex detection of BCR-ABL transcript variants and JAK2 V617F mutation simultaneously using the reverse transcriptase polymerase chain reaction (RT-PCR)-based reverse dot-blot hybridization (RDB) method. MATERIAL AND METHODS: Separate biotinylated RT-PCR primers were designed to amplify specific BCR-ABL transcripts and JAK2 V617F mutant alleles. Specific hybridization of RT-PCR products with arrays of membrane-bound probes followed by colorimetric development would allow simultaneous visualization of BCR-ABL and/or JAK2 mutant transcripts in a given specimen. To validate the RDB method, we used cDNA specimens previously referred to our laboratory for routine clinical testing of BCR-ABL and/or JAK2. RESULTS: The limit of detection or analytical sensitivity of the RDB method using cDNA specimens was 0.5% and 6.25% in detecting BCR-ABL and JAK2 mutant transcripts, respectively. The diagnostic specificity and sensitivity to detect BCR-ABL and JAK2 were 100% and 92.3% (N = 38); and 100% and 100% (N = 27), respectively. RDB also detected BCR-ABL transcripts in 22% of JAK2 V617F mutation-positive samples (N = 14). CONCLUSIONS: RT-PCR RDB is a promising qualitative multiplex method to detect BCR-ABL and JAK2 mutant transcripts simultaneously.