A novel real-time PCR assay for highly specific detection and quantification of vaginal lactobacilli.

Abstract:

:PCR detection and quantification of vaginal lactobacilli remains problematic because of the high level of genetic heterogeneity and taxonomic complexity within the genus Lactobacillus. The aim of the present study was to identify conserved sequences among the genomes of major species of vaginal lactobacilli that could be used for the development of a PCR-based method for quantitative determination of vaginal microbiota-specific lactobacilli. Comparative analysis of the genomes of several species of vaginal lactobacilli allowed us to identify conserved regions in the rplK gene, which encodes ribosomal protein L11, and to design group-specific PCR primers and a probe for selected species from the L. acidophilus complex, including major vaginal lactobacilli Lactobacillus crispatus, L. gasseri, L. iners and L. jensenii as well as other species that are less common in vaginal microbiota. The applicability of the new assay in routine diagnostic testing was evaluated using a set of clinical samples. The assay was able to detect and quantify vagina-associated lactobacilli within a wide range of initial DNA template concentrations, indicating promising potential for clinical applications.

journal_name

Mol Cell Probes

authors

Demkin VV,Koshechkin SI,Slesarev A

doi

10.1016/j.mcp.2016.11.006

subject

Has Abstract

pub_date

2017-04-01 00:00:00

pages

33-39

eissn

0890-8508

issn

1096-1194

pii

S0890-8508(16)30116-5

journal_volume

32

pub_type

杂志文章
  • A method for the rapid construction of cRNA standard curves in quantitative real-time reverse transcription polymerase chain reaction.

    abstract::Quantification of nucleic acids, especially of mRNA, is increasingly important in biomedical research. The recently developed quantitative real-time polymerase chain reaction (PCR) - a highly sensitive technology for the rapid, accurate and reproducible quantification of gene expression - offers major advantages over ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.2002.0405

    authors: Fronhoffs S,Totzke G,Stier S,Wernert N,Rothe M,Brüning T,Koch B,Sachinidis A,Vetter H,Ko Y

    更新日期:2002-04-01 00:00:00

  • miR-410-5p promotes the development of diabetic cardiomyopathy by suppressing PIM1-induced anti-apoptosis.

    abstract::Diabetic cardiomyopathy (DCM) is a common complication of diabetes mellitus that can cause many severe symptoms, such as heart failure, arrhythmia, and sudden death. However, the molecular mechanisms underlying cardiac dysfunction in DCM remain elusive. In this study, we found that miR-410-5p was increased in the myoc...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2020.101558

    authors: Xia X,Liang Y,Zheng W,Lin D,Sun S

    更新日期:2020-08-01 00:00:00

  • Detection of hepatitis B virus DNA in serum with nucleic acid probes labelled with 32P, biotin, alkaline phosphatase or sulphone.

    abstract::To facilitate the clinical application of dot-blot hybridization for assaying hepatitis B virus (HBV) DNA, we compared the ability of nucleic acid probes labelled with 32P or with various non-radioactive markers to detect HBV DNA in patient serum. Cloned HBV DNA was hybridized with (1) 32P-labelled HBV DNA cloned in M...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/0890-8508(91)90053-m

    authors: Valentine-Thon E,Steinmann J,Arnold W

    更新日期:1991-08-01 00:00:00

  • Specific detection of Angiostrongylus cantonensis in the snail Achatina fulica using a loop-mediated isothermal amplification (LAMP) assay.

    abstract::Angiostrongylus cantonensis, a rat lungworm, can cause eosinophilic meningitis and angiostrongyliasis in humans following ingestion of contaminated foods or intermediate/paratenic hosts with infective larvae. The snail Achatina fulica is one of the important intermediate hosts of A. cantonensis and is commonly eaten b...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2011.04.002

    authors: Liu CY,Song HQ,Zhang RL,Chen MX,Xu MJ,Ai L,Chen XG,Zhan XM,Liang SH,Yuan ZG,Lin RQ,Zhu XQ

    更新日期:2011-08-01 00:00:00

  • Mutation screening of the chromosome 8q24.3-human activity-regulated cytoskeleton-associated gene (ARC) in idiopathic generalized epilepsy.

    abstract::Idiopathic generalized epilepsy (IGE) comprises a heterogeneous group of disorders, in which a high genetic predisposition and a complex mode of inheritance have been suggested. However, genes, which confer liability to common IGE subtypes including juvenile myoclonic epilepsy (JME) and childhood absence epilepsy (CAE...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.2000.0314

    authors: Haug K,Kremerskothen J,Hallmann K,Sander T,Dullinger J,Rau B,Beyenburg S,Lentze MJ,Barnekow A,Elger CE,Propping P,Heils A

    更新日期:2000-08-01 00:00:00

  • A multiplex PCR for detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in clinical specimens.

    abstract::A multiplex PCR was developed that is capable of detecting four of the most important bacterial agents of atypical pneumonia, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in uncultured patient specimens. These organisms cause similar symptomologies and are often not...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2005.05.002

    authors: McDonough EA,Barrozo CP,Russell KL,Metzgar D

    更新日期:2005-10-01 00:00:00

  • Endostar regulates EMT, migration and invasion of lung cancer cells through the HGF-Met pathway.

    abstract:AIM:Though Endostar (ES) could inhibit tumor growth by inhibiting tumor angiogenesis, other possible mechanisms have been less reported. This study aims to investigate the role of ES in the treatment of lung cancer from the perspective of macrophage-mediated epithelial mesenchymal transformation (EMT). METHODS:THP1 ce...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2019.05.003

    authors: Shen Y,Chen Q,Li L

    更新日期:2019-06-01 00:00:00

  • Identification of six novel CFTR mutations in a sample of Italian cystic fibrosis patients.

    abstract::The spectrum of cystic fibrosis (CF) mutations has been determined in many populations of different ethnic and geographic origins. However, in the south of Europe, the commonest mutation, delta F508, accounts for only about 50% of CF chromosomes, while identification of most of the other mutant alleles has not been ac...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/s0890-8508(95)80038-7

    authors: Férec C,Novelli G,Verlingue C,Quéré I,Dallapiccola B,Audrézet MP,Mercier B

    更新日期:1995-04-01 00:00:00

  • Use of antibodies against the P36 protein of Mycoplasma hyopneumoniae for the identification of M. hyopneumoniae strains.

    abstract::Mycoplasma hyopneumoniae, the principal aetiological agent of porcine enzootic pneumonia, synthesizes a 36 kDa protein (P36) which is an early and strong immunogenic factor in experimentally and naturally infected swine. Polyclonal antibodies were made against the recombinant P36 protein in rabbits and used for the id...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/s0890-8508(05)80017-9

    authors: Stipkovits L,Nicolet J,Haldimann A,Frey J

    更新日期:1991-12-01 00:00:00

  • The construction and use of a PCR internal control.

    abstract::An example of the application and contruction of a polymerase chain reaction (PCR) internal control is presented. The internal control is synthesized in one PCR reaction. The primers used in this reaction possess 5' over-hanging ends which are identical to the primers used in the diagnostic reaction, whereas their 3' ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.1998.0170

    authors: Sachadyn P,Kur J

    更新日期:1998-10-01 00:00:00

  • Experiences on the application of the polymerase chain reaction in a diagnostic laboratory.

    abstract::Double polymerase chain reaction (PCR) assays with nested primers have been applied in a routine laboratory for the diagnosis of herpes-, pesti- and retroviral infections of animals. Various methods and tools have been tested to prevent and to eliminate false positive results as well as to visualize the PCR products (...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,评审

    doi:10.1006/mcpr.1993.1035

    authors: Belák S,Ballagi-Pordány A

    更新日期:1993-06-01 00:00:00

  • Development and validation of a cost-effective in-house method, tetra-primer ARMS PCR assay, in genotyping of seven clinically important point mutations.

    abstract::The single nucleotide polymorphism (SNP) genotyping is currently considered as a particularly valuable tool for the diagnosis of different pathologies. For this reason, over the past several years a great deal of effort has been devoted to developing accurate, rapid, and cost-effective technologies for SNP analysis. A...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2011.04.005

    authors: Etlik O,Koksal V,Arican-Baris ST,Baris I

    更新日期:2011-08-01 00:00:00

  • A novel technique for rapid automated genotyping of DNA polymorphisms in the mouse.

    abstract::The ability to rapidly and reliably genotype mice is an important concern. Traditional methods employ labour intensive and time consuming techniques such as test crossing, gel electrophoresis or nucleic acid hybridization. Here we show that a new molecular biology workstation, the WAVE DNA Fragment Analysis System, ca...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.1999.0239

    authors: Kuklin A,Davis AP,Hecker KH,Gjerde DT,Taylor PD

    更新日期:1999-06-01 00:00:00

  • Rapid diagnosis of spinal muscular atrophy using tetra-primer ARMS PCR assay: simultaneous detection of SMN1 and SMN2 deletion.

    abstract::Spinal muscular atrophy (SMA), the leading genetic cause of death in childhood, is an autosomal recessive neuromuscular disorder characterized by progressive muscle weakness, associated with deletions of the survival motor neuron 1 (SMN1) gene. Approximately 94% of SMA patients carry homologous deletions of SMN1 exon(...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2009.12.001

    authors: Baris I,Etlik O,Koksal V,Arican-Baris ST

    更新日期:2010-06-01 00:00:00

  • Successful quantification of cytomegalovirus DNA by competitive PCR and detection with capillary electrophoresis.

    abstract::Human cytomegalovirus (HCMV) is responsible for severe infections in immunocompromised patients. Viral load has recently been identified as one of the major risk factors for subsequent development of HCMV disease. In this context, we developed a protocol allowing rapid, sensitive and precise quantification of HCMV DNA...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.1996.0071

    authors: Poirier-Toulemonde AS,Imbert-Marcille BM,Ferré-Aubineau V,Besse B,Le Roux MG,Cantarovich D,Billaudel S

    更新日期:1997-02-01 00:00:00

  • Twelve hour real-time PCR technique for the sensitive and specific detection of Salmonella in raw and ready-to-eat meat products.

    abstract::Rapid pathogen testing is vital to the food industry. Enzyme immunoassays (EIA) provide reliable negative results in 48 h, but a presumptive positive (suspect) EIA result must be confirmed by traditional culture methods, requiring an additional 72 h. Polymerase chain reaction (PCR) testing technology is accepted as an...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2003.09.007

    authors: Ellingson JL,Anderson JL,Carlson SA,Sharma VK

    更新日期:2004-02-01 00:00:00

  • Detection and molecular typing of Campylobacter jejuni in fecal samples by polymerase chain reaction.

    abstract::In order to determine whether polymerase chain reaction (PCR) could be used to detect Campylobacter jejuni directly in stool samples, DNA from 66 frozen culture positive and negative fecal samples was purified by column chromatography. The flaA gene was amplified using primers directed against the conserved 5' and 3' ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.1996.0011

    authors: Waegel A,Nachamkin I

    更新日期:1996-04-01 00:00:00

  • Fast and sensitive quantitative detection of HIV DNA in whole blood leucocytes by SYBR green I real-time PCR assay.

    abstract::The aim of this study was the development of a real-time PCR for HIV DNA quantification in whole blood leucocytes providing an alternative assay to those already described, almost based on the gag gene detection. The technique (pbs-rtPCR assay) is more rapid (the whole assay required less than 5h), easy to perform, om...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2007.05.005

    authors: Casabianca A,Gori C,Orlandi C,Forbici F,Federico Perno C,Magnani M

    更新日期:2007-10-01 00:00:00

  • Two-color multiplex assay for the identification of orthopox viruses with real-time LUX- PCR.

    abstract::The LUX [Light Upon eXtension] is a real-time detection system that can be used for the detection and quantification of pathogens nucleic acids. In this study we used a universal LUX approach, a variation of the LUX detection system, for identifying Orthopoxvirus nucleic acids in real time. This approach enables the d...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2005.05.003

    authors: Aitichou M,Javorschi S,Ibrahim MS

    更新日期:2005-10-01 00:00:00

  • Characterization of a Caenorhabditis elegans glc seven-like phosphatase (gsp) orthologue from Haemonchus contortus (Nematoda).

    abstract::A full-length complementary DNA (cDNA; designated Hc-stp-1) encoding a serine/threonine phosphatase (Hc-STP-1) was isolated from Haemonchus contortus, a strongylid nematode parasite of small ruminants. Hc-stp-1 was shown to be transcribed in males of both adults and fourth-stage larvae, but not in females, early larva...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2010.02.001

    authors: Campbell BE,Rabelo EM,Hofmann A,Hu M,Gasser RB

    更新日期:2010-08-01 00:00:00

  • Development of a p28-based PCR assay for Ehrlichia chaffeensis.

    abstract::Detection of Ehrlichia chaffeensis is necessary to study interactions between the parasite and its vertebrate and invertebrate hosts. The purpose of this study was to develop a sensitive, specific PCR assay for E. chaffeensis based on the outer membrane protein gene, p28. Candidate primer sets were identified and rank...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2003.10.002

    authors: Wagner ER,Bremer WG,Rikihisa Y,Ewing SA,Needham GR,Unver A,Wang X,Stich RW

    更新日期:2004-04-01 00:00:00

  • Exploring transcriptional conservation between Ancylostoma caninum and Haemonchus contortus by oligonucleotide microarray and bioinformatic analyses.

    abstract::In this study, we identified, using an established oligonucleotide microarray platform for the parasitic nematode Haemonchus contortus, transcripts that are 'conserved' between serum-activated and non-activated L3s of Ancylostoma caninum (aL3 and L3, respectively) and H. contortus by cross-species hybridization (CSH) ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2008.09.004

    authors: Cantacessi C,Loukas A,Campbell BE,Mulvenna J,Ong EK,Zhong W,Sternberg PW,Otranto D,Gasser RB

    更新日期:2009-02-01 00:00:00

  • Simultaneous detection of two cystic fibrosis alleles using dual-label time-resolved fluorometry.

    abstract::A simple dual-label hybridization test for normal and mutant cystic fibrosis (CF) alleles is described. The assay is based on time-resolved fluorometry (TRF), which allows the simultaneous detection of DNA probes labelled with different lanthanides from one hybridization reaction. DNA was liberated from dried blood di...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/0890-8508(92)90047-2

    authors: Iitiä A,Liukkonen L,Siitari H

    更新日期:1992-12-01 00:00:00

  • Direct extraction and molecular characterization of enteroviruses genomes from human faecal samples.

    abstract::Routine diagnosis of acute flaccid paralysis (AFP) is still based on classical virological procedures. Several enteroviruses serotypes are not easily isolated in cell cultures system used and routinely more than one passage in cell culture is performed. A total of 54 archived faecal samples were examined. The heteroge...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2007.12.001

    authors: Bolanaki E,Kottaridi C,Dedepsidis E,Kyriakopoulou Z,Pliaka V,Pratti A,Levidiotou-Stefanou S,Markoulatos P

    更新日期:2008-06-01 00:00:00

  • Molecular detection and identification of enteroviruses in children admitted to a university hospital in Greece.

    abstract::Although enteroviral infections occur frequently during childhood, the circulation of particular serotypes has never been studied in Greece. The objectives of the present report were molecular detection and identification of human enteroviruses in children admitted with nonspecific febrile illness or meningitis to a u...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2011.06.001

    authors: Siafakas N,Attilakos A,Vourli S,Stefos E,Meletiadis J,Nikolaidou P,Zerva L

    更新日期:2011-10-01 00:00:00

  • Multiplex PCR detection of Campylobacter jejuni and Arcobacter butzleri in food products.

    abstract::Arcobacter is a recently described species, previously considered part of the Campylobacter family. A sensitive assay such as that provided by PCR could help to distinguish the closely related Arcobacter from Campylobacter. A PCR method to specifically detect both Campylobacter jejuni and Arcobacter butzleri in the sa...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.2000.0290

    authors: Winters DK,Slavik MF

    更新日期:2000-04-01 00:00:00

  • PVT1 induces NSCLC cell migration and invasion by regulating IL-6 via sponging miR-760.

    abstract::Non-small-cell lung carcinoma (NSCLC) accounts for approximately 80% of lung cancers with a high metastatic potential. Elucidating the mechanism of NSCLC metastasis will provide new promising targets for NSCLC therapy and benefit its prognosis. Plasmacytoma variant translocation 1 (PVT1) has been proven to be overexpr...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2020.101652

    authors: Su XH,Zhu YR,Hou YJ,Li K,Dong NH

    更新日期:2020-12-01 00:00:00

  • Development of a TB green II-based duplex real-time fluorescence quantitative PCR assay for the simultaneous detection of porcine circovirus 2 and 3.

    abstract::Porcine circovirus 3 (PCV3), as a newly emerged circovirus, is widely distributed in pig populations worldwide. Co-infection of PCV2 and PCV3 has been reported frequently in clinical samples. In the present study, a TB Green II-based duplex real-time polymerase chain reaction (qPCR) was developed to rapidly and differ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2019.04.001

    authors: Zhao Y,Han HY,Fan L,Tian RB,Cui JT,Li JY,Chen HY,Yang MF,Zheng LL

    更新日期:2019-06-01 00:00:00

  • A rapid one-tube PCR method for simultaneously differentiating homozygotes and heterozygotes of the Sp1 binding site polymorphism in collagen type Ialpha1.

    abstract::Rapid detection of single-base changes is fundamental to molecular medicine. PCR amplification of specific alleles (PASA) has previously been used as a rapid method of genotyping single-nucleotide changes, but one reaction is required for each allele. This paper describes a Bidirectional PASA (Bi-PASA) method, which w...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1016/j.mcp.2006.10.007

    authors: Lundberg GA

    更新日期:2007-06-01 00:00:00

  • HyBeacon probes: a new tool for DNA sequence detection and allele discrimination.

    abstract::Technologies that permit rapid investigation of DNA sequences, such as those containing single nucleotide polymorphisms (SNPs), are of great consequence to many sectors that perform molecular diagnostic analyses. We have developed a novel fluorescent oligonucleotide probe technology, termed HyBeacons, which provides a...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章

    doi:10.1006/mcpr.2001.0384

    authors: French DJ,Archard CL,Brown T,McDowell DG

    更新日期:2001-12-01 00:00:00