Comparative genotyping of Streptococcus mutans by repetitive extragenic palindromic polymerase chain reaction and multilocus sequence typing.
Comparative genotyping of Streptococcus mutans by repetitive extragenic palindromic polymerase chain reaction and multilocus sequence typing.
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通过重复外源回文聚合酶链反应和多位点序列分型对变形链球菌进行比较基因分型。
DOI：
10.1111/omi.12002
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发表时间：
2013
影响因子：
3.7
通讯作者：
Childers,NK
中科院分区：
文献类型：
--
作者：
Momeni,SS;Whiddon,J;Moser,SA;Cheon,K;Ruby,JD;Childers,NK
变形链球菌的遗传多样性已被广泛研究，使用各种基因分型方法。重复基因外回文聚合酶链反应（rep-PCR）是一种用于筛选大量细菌分离株的基因分型方法。这项由两部分组成的研究使用多位点序列分型（MLST）分析来评估先前使用rep-PCR鉴定为独特的基因型。在第一部分中，从代表8000株临床分离株的22个变异链球菌-PCR基因型组中各选择一株分离株。对于第二部分，从6个最常见的基因型组（GG）中选择4个额外的分离株进行进一步分析。使用8个管家突变链球菌基因座进行真实的实时PCR，并对扩增子进行测序。使用CLC DNAWorkbenchand进行序列数据分析，并使用中野方案将口腔链球菌的等位基因与PubMLST数据库进行比较。使用具有自举的最小进化方法，用MEGA评估串联序列。所有22个rep-PCR基因型通过MLST分析是独特的。在rep-PCR GG中，MLST在三个组中与rep-PCR匹配，证明了克隆性;三个组在MLST中表现出更大的多样性。三个克隆群的发现是这项研究的独特之处，表明变异链球菌的基因型在无关的受试者之间是共享的。此外，MLST定义了19个新的等位基因和26个新的序列类型，已被PubMLST确认和注册。简化了处理方法，并建议使用MLST和rep-PCR的过程。总之，MLST验证了rep-PCR是一种可靠且具有成本效益的方法，可用于筛选大量变形链球菌菌株进行流行病学研究。
The genetic diversity ofStreptococcus mutanshas been extensively studied using a variety of genotyping methods. Repetitive extragenic palindromic‐polymerase chain reaction (rep‐PCR) is a genotyping approach used for screening large numbers of bacterial isolates. This two‐part study used multilocus sequence typing (MLST) analysis to evaluate genotypes previously identified as unique using rep‐PCR. In part one, an isolate was selected from each of the 22S. mutansrep‐PCR genotype groups representing 8000 clinical isolates. For part two, four additional isolates were selected from the six most commonly occurring genotype groups (GG) for further analysis. Real‐time PCR was performed using eight housekeepingS. mutansgene loci and the amplicons were sequenced. Sequence data analysis was performed using CLC DNAWorkbenchand alleles were compared with the PubMLST database for Oral Streptococcus using the Nakano scheme. Concatenated sequences were evaluated with MEGA using a minimum evolution method with bootstrap. All 22 rep‐PCR genotypes were unique by MLST analysis. Within rep‐PCR GGs, MLST matched rep‐PCR in three groups demonstrating clonality; three groups exhibited more diversity with MLST. The discovery of three clonal groups is unique to this study and suggests thatS. mutansgenotypes are shared between unrelated subjects. Furthermore, MLST defined 19 new alleles and 26 new sequence types that have been confirmed and registered with PubMLST. Methods for processing were streamlined and a process for using MLST with rep‐PCR is suggested. In conclusion, MLST verified that rep‐PCR is a reliable and cost‐effective method for screening large numbers ofS. mutansstrains for epidemiological study.
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DOI：
--
发表时间：
1976
期刊：
影响因子：
--
作者：
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通讯作者：
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影响因子：
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4.8
作者：
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影响因子：
4.8
作者：
E. Mulvihill;R. Palmiter
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R. Palmiter
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DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
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作者：
Wang,SY;Gudas,LJ
通讯作者：
Gudas,LJ