Rapid and simple detection of <i>Ureaplasma</i> species from vaginal swab samples using a loop‐mediated isothermal amplification method

Fuwa, Kazumasa; Seki, Mitsuko; Hirata, Y.; Yanagihara, Itaru; Nakura, Yukiko; Takano, Chika; Kuroda, Kensuke; Hayakawa, Satoshi

doi:10.1111/aji.12771

Cited by 8 publications

(9 citation statements)

References 27 publications

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“…also require a complicated enrichment culture. In addition to these diagnostic modalities, we have constructed a rapid, sensitive and accurate loop‐mediated isothermal amplification method for ureaplasma detection . However, pregnant patients are not routinely screened for ureaplasma infection because nucleic acid detection of Ureaplasma spp.…”

Section: Therapeutic Options For Genital Ureaplasma Infection In Pregmentioning

confidence: 99%

Current understanding and treatment of intra‐amniotic infection with Ureaplasma spp.

Tantengco

Yanagihara

2019

J of Obstet and Gynaecol

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Considerable evidence has shown that intra-amniotic infection with Ureaplasma spp. increases the risk of chorioamnionitis and preterm labor. Ureaplasma spp. are among the smallest organisms, and their isolation is uncommon in routine clinical practice because of their size and high auxotrophy. Although Ureaplasma spp. have been reported as causative agents of preterm birth, they also have a high incidence in vaginal swabs collected from healthy reproductive-age women; this has led to questions on the virulence of Ureaplasma spp. and to them being considered as harmless commensal bacteria. Therefore, many efforts have been made to clarify the pathogenicity of Ureaplasma spp. at the molecular level. Ureaplasma spp. are surrounded by lipoproteins, including multiple-banded antigen. Both multiple-banded antigen and its derivative, that is, the synthetic lipopeptide, UPM-1, induce an inflammatory response in a preterm mice model, which was adequate to cause preterm birth or stillbirth. In this review, we present an overview of the virulence mechanisms of Ureaplasma spp. and treatment of ureaplasma infection during pregnancy to prevent possible serious sequelae in infants. In addition, relevant mechanisms underlying antibiotic resistance in Ureaplasma spp. are discussed. Ureaplasma spp. are naturally resistant against β-lactam antibiotics because of the lack of a cell wall. Azithromycin is one of the effective agents that can control intrauterine ureaplasma infection. In fact, macrolide-and fluoroquinolone-resistant isolates of Ureaplasma spp. have already been observed in perinatal practice in Japan.

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Section: Therapeutic Options For Genital Ureaplasma Infection In Pregmentioning

confidence: 99%

Current understanding and treatment of intra‐amniotic infection with Ureaplasma spp.

Tantengco

Yanagihara

2019

J of Obstet and Gynaecol

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“…The cloned fragments of the ureB gene of U. parvum (SV3F4) [16] and U. urealyticum (UU3) [17] were used as a positive control for each Ureaplasma species. The cloning procedure for ureB genes was performed in accordance with a previous report [15].…”

Section: Preparation For Positive Controlsmentioning

confidence: 99%

“…LAMP primers targeting the ureB gene for the detection of U. parvum or U. urealyticum were taken from the published study by Fuwa et al (S1 Table) [15]. LAMP reactions were performed in a 25-μl volume consisting of 5 μl template, 8 U of Bst DNA polymerase, 25 mM deoxynucleoside triphosphates, 4 M betaine, 1.5 M Tris-HCl (pH 8.8), 2.5 M KCL, 1 M (NH 4 )SO 4 , 1 M MgSO 4 , and 20% Tween 20.…”

Section: Lamp Reactionmentioning

confidence: 99%

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Validation of the loop-mediated isothermal amplification method for rapid and sensitive detection of Ureaplasma species in respiratory tracts of preterm infants

et al. 2021

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Introduction A simple and rapid diagnosis of Ureaplasma spp. is required for the choice of the appropriate antibiotic. However, an ideal detection method has not been available. This study examines the efficacy of the loop-mediated isothermal amplification (LAMP) assay, which provides rapid and sensitive results, to detect Ureaplasma spp. in respiratory tract samples of preterm infants. Methods The study included preterm infants born before 32 weeks of gestation admitted Kagoshima City Hospital from June 2018 to March 2020. Nasopharyngeal swabs and/or tracheal aspirates were obtained in the first seven postnatal days. One hundred sixty-seven nasopharyngeal swabs and 101 tracheal aspirates were analyzed by LAMP, culture, and quantitative real-time polymerase chain reaction. Results All 167 infants had a median (range) gestational age of 28.7 weeks (22.3–30.9) and birthweight 1030g (322–1828). One hundred sixty-seven nasopharyngeal swabs and 101 tracheal aspirates were obtained. In the results of nasopharyngeal swabs, the sensitivity and specificity of LAMP were 73.9% (17/23) and 97.2% (140/144), whereas those of quantitative real-time polymerase chain reaction were 73.9% (17/23) and 95.8% (138/144), compared to culture. In the results of tracheal aspirates, the sensitivity and specificity of LAMP were 89.5% (17/19) and 92.7% (76/82), whereas those of quantitative real-time polymerase chain reaction were 89.5% (17/19) and 93.9% (77/82), compared to culture. Conclusions The LAMP assay showed similar sensitivity and specificity with quantitative real-time polymerase chain reaction in the respiratory tracts of preterm infants including extremely preterm infants during the immediate postnatal period. Therefore, the LAMP is a practical alternative for the early detection so that appropriate antibiotics can be administered for preventing BPD.

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“…With more and more scientists focusing their attention on the application of LAMP technology, the range of its use is not limited to the bacteria detection and identification any more [14]. The LAMP-PCR was developed and employed to detect species that cause chorioamnionitis and premature labor, Ureaplasma parvum, and Ureaplasma urealyticum [15].…”

Section: Loop-mediated Isothermal Amplificationmentioning

confidence: 99%

PCR and Infectious Diseases

Zauli¹

2020

Synthetic Biology - New Interdisciplinary Science

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Since the 1950s, the medical community has been faced with infectious diseases, which have brought significant public health and financial challenges. Currently, routine testing for the laboratory diagnosis for infectious agents is based on cell culture, serological, and molecular methods. However, cell culture-based methods are used mainly in research laboratories and are less sensitive methods when compared with serological and molecular methods. The diagnosis of infectious diseases has been revolutionized by the development of molecular techniques, mainly with the applications of polymerase chain reaction (PCR). The high sensitivity, specificity, and ease with which the PCR can be used to detect genetic sequences known have led to your wide application in science. A great number of qualitative and quantitative molecular assays are mostly based on what have been described such as real-time PCR, multiplex PCR, LAMP-PCR, and digital PCR. These assays could identify active infection by detecting infectious agents and nucleic acid in various clinical conditions including arboviruses, sexually transmitted infections, and bacterial infections. Further advancement of molecular technology is needed to improve the capacity to detect infectious agents in order to control the spread of infectious diseases and lead to appropriate actions which help to benefit patients and health-care workers themselves.

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Rapid and simple detection of Ureaplasma species from vaginal swab samples using a loop‐mediated isothermal amplification method

Abstract: The LAMP detection method outperformed the culture and PCR methods. Early detection enables appropriate antibiotic selection for improved prenatal outcomes.

Cited by 8 publications

References 27 publications

Current understanding and treatment of intra‐amniotic infection with Ureaplasma spp.

Current understanding and treatment of intra‐amniotic infection with Ureaplasma spp.

Validation of the loop-mediated isothermal amplification method for rapid and sensitive detection of Ureaplasma species in respiratory tracts of preterm infants

PCR and Infectious Diseases

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