Sensitivity of the Standard <i>Chlamydia trachomatis</i> Culture Method Is Improved After One Additional In Vitro Passage

Song, Lili; Guo, Yuanli; Jiang, Yong; Liu, Yuanjun; Wang, Mei; Yu, Cong; Liu, Quanzhong

doi:10.1002/jcla.21924

Cited by 15 publications

(5 citation statements)

References 12 publications

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“…This sensitivity may be further reduced by culture overgrowth by commensal bacteria [47]. Recently, however, Shao et al demonstrated an increased culture sensitivity by performing additional in vitro passages compared to standard protocols and reported a culture sensitivity of 80% after three blind culture passages [48], but this will lengthen the time to diagnosis even more. Culture methods have mostly long turnaround times of at least 24 h and up to 3 days post infection without additional in vitro passages and with its labor intensive and demanding procedure, chlamydia culturing is not available in most routine laboratory settings.…”

Section: Strengths and Limitationsmentioning

confidence: 99%

Review ofChlamydia trachomatisviability methods: assessing the clinical diagnostic impact of NAAT positive results

Janssen

Dirks

Dukers-Muijrers

et al. 2018

Expert Review of Molecular Diagnostics

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Chlamydia trachomatis (chlamydia) is the most commonly diagnosed bacterial sexually transmitted infection (STI) worldwide. The advancement of molecular techniques has made chlamydia diagnostics infinitely easier. However, molecular techniques lack the information on chlamydia viability. Where in routine diagnostics the detection of chlamydia DNA or RNA might suffice, in other patient scenarios, information on the viability of chlamydia might be essential. Areas covered: In this review, the authors discuss the specific strengths and limitations of currently available methods to evaluate chlamydia viability: conventional cell culture, messenger RNA (mRNA) detection and viability-PCR (V-PCR). PubMed and Google Scholar were searched with the following terms: Chlamydia trachomatis, Treatment failure, Anal chlamydia, Microbial viability, Culture, Viability-PCR, Messenger RNA, and Molecular diagnostics Expert commentary: Several techniques are currently available to determine chlamydia viability and thus the clinical relevance of a positive test result in clinical samples. Depending on the underlying research question, all three discussed techniques have their merits when testing for viability. However, mRNA methods show the most promise in determining the presence of a true infection, in case the chlamydia reticulate body can be specifically detected. Further research is needed to understand how to best apply viability testing in current chlamydia diagnostics.

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Section: Strengths and Limitationsmentioning

confidence: 99%

Review ofChlamydia trachomatisviability methods: assessing the clinical diagnostic impact of NAAT positive results

Janssen

Dirks

Dukers-Muijrers

et al. 2018

Expert Review of Molecular Diagnostics

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“…It has been inferred that the accumulation of antibiotics in patients can limit CT growth during early passages. 34 In addition, some experts have reported that in vitro exposure to several β-lactam antibiotics causes the reticulate body of CT to convert to the aberrant reticulate body phenotype found in a persistent state. 25 Therefore, the administration of amoxicillin can cause CT to enter a persistent state.…”

Section: Discussionmentioning

confidence: 99%

“…A two-passage culture method was applied in all clinical swabs because we found that there were only a small amount of inclusions formed in the clinical swabs in the first passage; thus, the sensitivity of CT iodine staining could be increased with multiple passages. 34 The successfully cultured CT strains were stored in sucrose phosphate glutamate (218 mM sucrose, 3.76 mM KH 2 PO 4 , 7.1 mM K 2 HPO 4 , and 5 mM GlutaMAX-100—all purchased from the Solarbio Technological Co, Ltd. Beijing, China) storage medium at −80 °C for further use.…”

Section: Methodsmentioning

confidence: 99%

The Effect of Amoxicillin Pre-Exposure on Treatment Outcomes and Antimicrobial Susceptibility in Patients with Urogenital Chlamydia trachomatis Infection

You¹,

Liao²,

Wang³

et al. 2023

IDR

Self Cite

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Purpose We investigated the influence of amoxicillin pre-exposure on treatment outcomes, Chlamydia trachomatis (CT) culture, the presence of drug-resistant genes, minimum inhibitory concentrations (MICs), and fractional inhibitory concentrations (FICs) in CT clinical strains. Additionally, we explored the effect of different antimicrobial combinations on CT. Patients and Methods Clinical data of 62 patients with CT infection were recorded. Of these, 33 had pre-exposure to amoxicillin and 29 did not. Among patients with pre-exposure, 17 received azithromycin and 16 received minocycline. Among the patients without pre-exposure, 15 received azithromycin and 14 received minocycline. All patients underwent microbiological cure follow-ups one month after completing the treatment. 23S rRNA gene mutations, acquisition of tet (M) and tet (C) were detected using reverse transcription PCR (RT-PCR) and PCR, respectively. The MICs and FICs of azithromycin, minocycline, and moxifloxacin, alone or in combination, were determined using the microdilution and checkerboard methods, respectively. Results More cases of treatment failure occurred in pre-exposed patients, in both treatment groups ( P <0.05). No 23S rRNA gene mutations or tet (M) and tet (C) acquisitions were found. More inclusion bodies were cultured from patients without amoxicillin pre-exposure than from those with pre-exposure ( P <0.0001). The MICs of all antibiotics were higher in pre-exposed patients than in those without pre-exposure ( P <0.01). The FICs of azithromycin plus moxifloxacin were lower than those of the other antibiotic combinations ( P <0.0001). The synergy rate of azithromycin plus moxifloxacin was significantly higher than those of azithromycin plus minocycline and minocycline plus moxifloxacin ( P <0.001). The FICs of all antibiotic combinations were comparable between isolates from the two patient groups (all P >0.05). Conclusion Pre-exposure to amoxicillin in CT patients may inhibit CT growth and decrease sensitivity of CT strains to antibiotics. Azithromycin plus moxifloxacin may be a promising treatment regimen for genital CT infections with treatment failure.

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“…Altogether, these approaches will break the transmission chain and prevent serious sequels. Regarding the detection methods, Chlamydia is not effectively detected by the conventional methods (using culture) because, despite the associated higher specificity, its sensitivity has a range from 3.9 to 80%, which is related to false negatives (~39%) [ 22 ]. Moreover, using cultures is expensive, requires approximately three to seven days, and is not technically simple.…”

Section: Introductionmentioning

confidence: 99%

Chlamydia trachomatis as a Current Health Problem: Challenges and Opportunities

Rodrigues

Sousa²,

Vale

2022

Diagnostics

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Chlamydia is one of the most common sexually transmitted bacterial infections (STIs) worldwide. It is caused by Chlamydia trachomatis (CT), which is an obligate intracellular bacterium. In some cases, it can occur in coinfection with other parasites, increasing the pathologic potential of the infection. The treatment is based on antibiotic prescription; notwithstanding, the infection is mostly asymptomatic, which increases the risk of transmission. Therefore, some countries have implemented Chlamydia Screening Programs in order to detect undiagnosed infections. However, in Portugal, there is no CT screening plan within the National Health Service. There is no awareness in the general healthcare about the true magnitude of this issue because most of the methods used are not Nucleic Acid Amplification Technology-based and, therefore, lack sensitivity, resulting in underreporting infection cases. CT infections are also associated with possible long-term severe injuries. In detail, persistent infection triggers an inflammatory milieu and can be related to severe sequels, such as infertility. This infection could also trigger gynecologic tumors in women, evidencing the urgent need for cost-effective screening programs worldwide in order to detect and treat these individuals adequately. In this review, we have focused on the success of an implemented screening program that has been reported in the literature, the efforts made concerning the vaccine discovery, and what is known regarding CT infection. This review supports the need for further fundamental studies in this area in order to eradicate this infection and we also suggest the implementation of a Chlamydia Screening Program in Portugal.

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Sensitivity of the Standard Chlamydia trachomatis Culture Method Is Improved After One Additional In Vitro Passage

Abstract: The results of this study support the use of a three-passage Chlamydia culture procedure to increase the detection sensitivity of this method.

Cited by 15 publications

References 12 publications

Review ofChlamydia trachomatisviability methods: assessing the clinical diagnostic impact of NAAT positive results

Review ofChlamydia trachomatisviability methods: assessing the clinical diagnostic impact of NAAT positive results

The Effect of Amoxicillin Pre-Exposure on Treatment Outcomes and Antimicrobial Susceptibility in Patients with Urogenital Chlamydia trachomatis Infection

Chlamydia trachomatis as a Current Health Problem: Challenges and Opportunities

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