Herrera-Mejía Julián, Campos-Vega Rocío, Wall-Medrano Abraham, Jiménez-Vega Florinda
Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo Envolvente del PRONAF y Estocolmo s/n, Ciudad Juárez 32310, Chihuahua, Mexico.
Programa de Posgrado en Alimentos del Centro de la República (PROPAC), Research and Graduate Studies in Food Science, School of Chemistry, Universidad Autónoma de Querétaro, Santiago de Querétaro 76010, Querétaro, Mexico.
Diagnostics (Basel). 2023 Jul 17;13(14):2387. doi: 10.3390/diagnostics13142387.
Many neurological disorders have a distinctive colonic microbiome (CM) signature. Particularly, children with autism spectrum disorders (ASD) exhibit a very dissimilar CM when compared to neurotypical (NT) ones, mostly at the species level. Thus far, knowledge on this matter comes from high-throughput (yet very expensive and time-consuming) analytical platforms, such as massive high-throughput sequencing of bacterial 16S rRNA. Here, pure (260/280 nm, ~1.85) stool DNA samples (200 ng.µL) from 48 participants [39 ASD, 9 NT; 3-13 y] were used to amplify four candidate differential CM markers [ (BF), (FP), (DV), (AM)], using micro-organism-specific oligonucleotide primers [265 bp (BF), 198 bp (FP), 196 bp (DV), 327 bp (AM)] and a standardized two-step [low (step 1: °-5 °C) to high (stage 2: °-0 °C) astringent annealing] PCR protocol (2S-PCR). The method was sensitive enough to differentiate all CM biomarkers in the studied stool donors [↑ abundance: NT (BF, FP, AM), ASD (DV)], and phylogenetic analysis confirmed the primers' specificity.
许多神经系统疾病都有独特的结肠微生物群（CM）特征。特别是，与神经典型（NT）儿童相比，自闭症谱系障碍（ASD）儿童的CM非常不同，主要在物种水平上。到目前为止，关于这一问题的知识来自高通量（但非常昂贵且耗时）的分析平台，如细菌16S rRNA的大规模高通量测序。在这里，使用来自48名参与者[39名ASD，9名NT；3 - 13岁]的纯（260/280 nm，~1.85）粪便DNA样本（200 ng.µL），使用微生物特异性寡核苷酸引物[265 bp（BF）、198 bp（FP）、196 bp（DV）、327 bp（AM）]和标准化的两步[低（步骤1：° - 5°C）到高（阶段2：° - 0°C）收敛退火]PCR方案（2S-PCR）扩增四个候选差异CM标记物[（BF）、（FP）、（DV）、（AM）]。该方法足够灵敏，能够区分所研究粪便供体中的所有CM生物标志物[丰度增加：NT（BF、FP、AM），ASD（DV）]，系统发育分析证实了引物的特异性。
