The amplification refractory mutation system (ARMS): A rapid and direct prenatal diagnostic technique for β-thalassaemia in Singapore

β-Thalassaemia major patients have chronic anaemia and since 3–4 per cent of Singaporeans carry the β-gene, prenatal diagnosis is essential. We evaluated the amplification refractory mutation system (ARMS) technique as a routine test for prenatal diagnosis of β-major. Six mutations along the β-gene were studied—41–42 (-TCTT), IVSII #654 (C-T), 17β (A-T), – 28 TATA (A-G), IVSI #5 (G-C), and IVSI #1 (G-T). Our results indicate that prenatal diagnosis using these mutations can be offered to 90 per cent (35/39) of our Chinese couples and 54·6 per cent (12/22) of our Malay couples at risk. Confirmation of ARMS results was carried out using allele-specific oligonucleotide hybridization. Prenatal diagnosis using ARMS was successfully carried out in nine cases which included a set of triplets and twins. The triplets were diagnosed with the β-trait carrying the 41–42 mutation. The couple with twins possessed the #654 mutation and one twin was diagnosed with the β-trait and the other with #654 homozygosity. Genomic sequencing of the undefined mutations in the Chinese couples revealed rarer mutations at − 29 and an ATG-AGG base substitution at the initiation codon for translation. In the Malay couples, genomic sequencing detected mutations at codon 15 (TGG-TAG) and codon 26 (GAG-AAG). We conclude that ARMS with its direct detection of amplified products by gel electrophoresis provides an accurate, rapid, and simpler method for our β-thalassaemia prenatal diagnosis programme in Singapore.     

水解酸化-序批式活性污泥法处理季戊四醇有机废水

在实验室研究的基础上 ,将水解酸化 序批式活性污泥 (HA SBR)处理工艺应用于季戊四醇生产废水治理工程中 ,运行结果表明 :CODCr去除率在 90 %以上 ,BOD5去除率在 85 %以上 ,所排废水各项指标达到国家排放标准。     

硝基苯降解菌在厌氧序批式反应器中处理硝基苯废水的应用

通过接种硝基苯降解菌于厌氧序批式反应器（ASBR），研究静态试验条件下和厌氧序批式工艺对硝基苯的降解规律，研究表明，应用生物强化技术处理硝基苯废水，具有很高的处理效率。     

利用混合菌群活性污泥法实现生物可降解塑料PHA的合成

PHA是一种生物可降解塑料，可从污水有机物中合成。文章在简要说明现阶段工业生产PHA情况的基础上，总结了利用混合菌种活性污泥法从污水有机物中合成PHA的优点。重点对两种从污水中生产PHA工艺——厌氧-好氧活性污泥工艺和好氧瞬时供料工艺进行了介绍，并给出了相应工艺流程。通过使用廉价的有机底物可以使PHA的生产价格降低到原来的一半，大约为每公斤4欧元左右。文章还讨论了控制工艺运行条件对PHA合成的重要性，并说明了不同底物组成对合成PHA性质的影响。     

水解酸化-序批式活性污泥法(ABR-SBR)处理电泳废水的研究

利用水解酸化 (采用缺氧折流板ABR反应器 ) -序批式活性污泥法 (SBR)工艺对汽车电泳涂装工艺废水进行实验研究 ,分析了ABR的反应时间、容积负荷、内置填料对废水可生化性改善的影响及SBR的运行方式、进水 pH、污泥负荷对有机物去除效果的影响     

两次污泥颗粒化过程中微生物群落的动态变化

利用序批式反应器(sequencing batch reactor,SBR)培养好氧颗粒污泥(aerobic granular sludge,AGS),在此期间发生了AGS破碎现象,后经培养,破碎污泥再次变为成熟的AGS.因此,采用Illumina Mi Seq PE300高通量测序技术研究了两次污泥颗粒化过程中微生物群落结构变化的差异,以期揭示有利于AGS形成的优势菌属;此外,利用实时定量聚合酶链式反应(quantitative polymerase chain reaction,q PCR)探究了两次污泥颗粒化过程中硝化微生物的动态变化.结果表明,在两次污泥颗粒化过程中,胞外蛋白质和胞外多糖的含量均增加;氨氧化古菌(ammonia oxidizing archaea,AOA)在第一次污泥颗粒化过程以及AGS成熟过程丰度增加,氨氧化细菌(ammonia oxidizing bacteria,AOB)虽然在第一次污泥颗粒化过程中丰度降低,但是在AGS培养过程中其丰度都一直高于AOA;微生物群落多样性随着AGS的形成而降低;变形菌门(Proteobacteria)相对丰度明显增加,分别增加了12.29%和5.90%;某些属于变形菌门的属其相对丰度也增加,其中,Candidatus Competibacter在两次污泥颗粒化过程中相对丰度增加最明显,并且在成熟的AGS中呈现很高的相对丰度,达到14.20%.总的来说,胞外蛋白质和胞外多糖含量的增加,可能促进了污泥颗粒化;AOA和AOB可能共同参与了AGS的氨氧化作用;Ca.Competibacter的富集可能有利于AGS的形成.     

Single-cell sequencing and mini-sequencing for preimplantation genetic diagnosis

There is increasing interest in the use of preimplantation genetic diagnosis (PGD) as an alternative to routine prenatal diagnosis. However, the costs associated with development and testing of new PGD protocols have forced some PGD centres to limit the number of diseases for which PGD is offered. One of the main factors in the design of new protocols, which affects cost and accuracy, is the choice of the mutation-detection technique. We have assessed the reliability of DNA sequencing and mini-sequencing for clinical diagnosis at the single-cell level and have found them to be rapid and accurate. Extensive optimisation for individual mutations is not usually necessary when employing these versatile techniques and consequently a smaller investment of time and resources should be required during development of new protocols. Additionally, we report single-cell protocols for the diagnoses of cystic fibrosis, sickle cell anaemia and β-thalassaemia, which utilise mini-sequencing. Unlike most mutation-detection techniques, mini-sequencing permits analysis of very small DNA fragments. Small amplicons experience low allele dropout (ADO) rates, and consequently this approach could potentially improve the reliability of PGD. Copyright © 2003 John Wiley & Sons, Ltd.     

低浓度铬对SBR中微生物抑制影响研究

研究了低浓度铬(三价和六价)对SBR生物系统的抑制影响,考察了两种不同SBR工艺(传统工艺和分段进水工艺)在处理含低浓度铬废水过程中常规的出水水质和活性污泥性状的变化,以及微生物群落的变迁.研究结果表明,在进水总铬(Cr(III)∶Cr(VI)=4∶1)浓度为0.5mg·L-1的条件下,传统工艺和分段进水工艺的氨氮去除率由99%分别下降至70%和65%,同时,磷酸盐去除率也由99%分别下降至51%和43%.当进水中总铬浓度达到1 mg·L-1时,分段进水工艺SBR系统的氨氮和磷酸盐去除率最终分别下降至44%和37%.此外,多糖和蛋白质的分泌量变化分别呈下降和上升趋势.高通量测序结果表明,活性污泥细菌群落丰富度和多样性受到了铬离子的影响,并且Nitrospira、Acidobacteria、Planctomycetes、Cyanobacteria和Candidatus_Accumulibacter等脱氮除磷功能菌种的生长都受到了一定程度的抑制,也与被抑制的SBR系统脱氮除磷去除率下降的宏观现象相吻合.     

SBR法处理高盐肝素废水运行参数优化研究

采用SBR法处理高盐肝素钠生产废水,探讨了曝气时间、进水p H、温度等参数对COD和氨氮降解效果及污泥生长的影响。结果表明,控制曝气时间为10 h、进水p H为7.5、反应温度在26~29℃时,COD和氨氮去除率均能稳定在85%以上;对氨氮去除效果较差的反应器,适当添加K+、Ca2+、Mg2+可以取得一定的改善作用,其中K+对Na+和NH4+双重毒性抑制的拮抗效应最明显;经参数优化后的SBR反应器对不同污泥负荷和盐度负荷具有较好的适应能力,受到冲击后COD和氨氮去除率均能在2~3个运行周期内恢复至90%以上。     

温度对厌氧环境下污泥中抗生素抗性基因行为特征的影响

抗性基因的转移传播使得污水处理厂成为其重要的储存库,对人类健康存在潜在风险。实验采用序批式厌氧反应器,温度设定分别为15℃、中温(30℃和36℃)、高温(50℃和60℃),探究温度对污泥厌氧条件下8种抗生素去除效果的影响,以及四环素类抗性基因(tet A、tet G、tet L、tet M、tet O、tet Q、tet W、tet X)、磺胺类抗性基因(sul I、sul II)和Ⅰ类整合子整合酶基因(int I 1)的行为特征。研究发现温度升高有利于抗生素及抗性基因的去除,15℃、中温和高温下总抗生素的平均去除率分别为45%、59%和78%;15℃、中温和高温下四环素类抗性基因分别削减0.52 log、0.90 log和1.50 log,磺胺类抗性基因分别削减0.56log、0.78 log和1.31 log。相关性分析发现,总抗性基因与总氮、氨氮、SCOD(溶解性COD)均存在显著相关性(R2=0.744、0.760、0.315,P0.05),而与总磷无显著相关性(P0.05)。int I 1与总氮、氨氮、SCOD皆存在显著相关性(R2=0.698、0.795、0.269,P0.05),而与总磷无显著相关性(P0.05)。说明微生物生长环境中的营养元素一定程度上影响着抗性基因的传播和扩散。