DETECCIÓN DE CHLAMYDIA TRACHOMATIS POR PCR EN TIEMPO REAL, EN RECIÉN NACIDOS EN EL HOSPITAL ISIDRO AYORA, QUITO-ECUADOR.

Introducción: La transmisión vertical de Chlamydia trachomatis se presenta del 60% -70% en los hijos de madres infectadas y el riesgo de adquirir conjuntivitis por Chlamydia trachomatis en estos neonatos varía entre el 18% al 70%. Objetivo: detectar la Chlamydia trachomatis por medio de qPCR (Reacción en Cadena de la Polimerasa por Tiempo Real) en los sacos conjuntivales en recién nacidos por parto normal. Métodos: Se realizó un estudio no experimental, transversal, analizando muestras procedentes de 178 niños nacidos vivos, en el Hospital Gíneco-obstétrico Isidro Ayora, se determinó la presencia de Chlamydia trachomatis en muestras de saco conjuntival, por medio de la reacción en cadena de la polimerasa los laboratorios de Biología Molecular de la Pontificia Universidad Católica del Ecuador. Resultados: la edad gestacional de los neonatos más frecuente es a término 75.29% que corresponde a 124 pacientes, resultados positivos para Chlamydia trachomatis por qPCR fueron 19 que corresponde al 10.7% y los resultados negativos 159 que corresponde al 89.3% de pacientes, el grupo de edad materna de los neonatos con resultados positivos para Chlamydia trachomatis por qPCR, más frecuente es el de 14 -19 años con 31.5%. Conclusiones: La conjuntivitis neonatal por Chlamydia trachomatis es una entidad de subdiagnósticada, que no responde al tratamiento con antibióticos tópicos habituales, donde la amplificación de ácidos nucleicos por medio de la qPCR ha sido un importante avance en el diagnóstico, debido a que la amplificación es extremadamente sensible (capaz de detectar una sola copia genómica) y altamente específica, inclusive en pacientes asintomáticos. Abstract Introduction: The vertical transmission of Chlamydia trachomatis occurs in 60% -70% in the children of infected mothers and the risk of contracting Chlamydia trachomatis conjunctivitis in these affected neonates between 18% and 70%. Objective: to detect Chlamydia trachomatis by means of qPCR (Real Time Polymerase Chain Reaction) in the conjunctival sacs in newborns by normal delivery. Methods: A non-experimental, cross-sectional study was carried out, analyzing analyzed samples of 178 live-born children, at the Gyneco-obstetric Hospital Isidro Ayora, the presence of Chlamydia trachomatis in conjunctival sac samples was determined by means of the chain reaction of polymerase molecular biology laboratories of the Pontifical Catholic University of Ecuador. Results: the most frequent gestational age of the neonates is at term 75.29% corresponding to 124 patients, positive results for Chlamydia trachomatis by qPCR were 19 corresponding to 10.7% and negative results 159 corresponding to 89.3% of patients, the group of maternal age of neonates with positive results for Chlamydia trachomatis by qPCR, more frequent is that of 14-19 years with 31.5%. Conclusions: Chlamydia trachomatis neonatal conjunctivitis is an underdiagnosed entity, which does not respond to treatment with usual topical antibiotics, where nucleic acid amplification by means of qPCR has been an important advance in diagnosis, due to extremely sensitive amplification. (capable of detecting a single genomic copy) and highly specific, including in asymptomatic patients

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West Nile Virus Chorioretinitis in the Presence of Negative Cerebrospinal Fluid Polymerase Chain Reaction Results

Journal of VitreoRetinal Diseases ◽

10.1177/2474126420979254 ◽

2021 ◽

pp. 247412642097925

Author(s):

Kareem Moussa ◽

Karen W. Jeng-Miller ◽

Leo A. Kim ◽

Dean Eliott

Keyword(s):

Cerebrospinal Fluid ◽

West Nile Virus ◽

Cross Sectional Study ◽

Nucleic Acid Amplification ◽

Sectional Study ◽

Chain Reaction ◽

West Nile ◽

Cross Sectional ◽

Positive Serum ◽

Polymerase Chain

Purpose: This work aims to evaluate the utility of nucleic acid amplification testing (NAAT) and serology in confirming West Nile Virus (WNV) infection in patients with suspected WNV chorioretinitis. Methods: A retrospective cross-sectional study was conducted of a cluster of patients who presented to the Retina Service of Massachusetts Eye and Ear between September and October 2018. Results: Three patients were identified with classic WNV chorioretinitis lesions with negative cerebrospinal fluid NAAT and positive serum serology findings. The diagnosis of WNV chorioretinitis was made based on the appearance of the fundus lesions and the presence of characteristic findings on fluorescein angiography as previously described in the literature. Conclusions: This report highlights 3 unique cases of WNV chorioretinitis in which NAAT of cerebrospinal fluid failed to identify WNV as the inciting agent. These cases stress the importance of serum serologic testing in diagnosing WNV infection.

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Assessment of rectal Chlamydia trachomatis viable load in women by viability-PCR

Sexually Transmitted Infections ◽

10.1136/sextrans-2019-054002 ◽

2019 ◽

Vol 96 (2) ◽

pp. 85-88 ◽

Cited By ~ 3

Author(s):

Kevin J H Janssen ◽

Petra Wolffs ◽

Mayk Lucchesi ◽

Nicole H T M Dukers-Muijrers ◽

Christian J P A Hoebe

Keyword(s):

Chlamydia Trachomatis ◽

Nucleic Acid Amplification ◽

Test Results ◽

Cross Sectional ◽

Convenience Sample ◽

Public Health Implication ◽

Significant Difference ◽

Rectal Swabs ◽

The Mean ◽

Positive Results

ObjectivesIn recent years, studies have demonstrated frequent rectal Chlamydia trachomatis (CT) detection in women, irrespective of reported anal sex or rectal symptoms. However, the clinical relevance and public health implication of rectal CT detection in women remain under debate. Therefore, evaluating CT viability may provide more insight into the relevance of standard routine nucleic acid amplification test (NAAT)–positive results.MethodsIn this cross-sectional explorative study, a convenience sample of female patients at our STI clinic aged 18 years or older, diagnosed with vaginal and/or rectal CT, were invited to participate. On return for treatment, rectal CT-diagnosed women were instructed to self-collect rectal swab samples before being treated. Standard COBAS 4800 CT/NG routine NAAT testing was applied for CT diagnosis. Rectal viable CT load was evaluated by using viability-PCR (V-PCR).Results53 women with rectal CT were included in this study; 86.8% (46/53) had a quantifiable rectal total CT load. Of women with quantifiable samples, 52.2% (24/46) had viable CT detected from rectal swabs by V-PCR, with a mean rectal viable CT load of 3.31 log10 CT/mL (range 1.16–6.22). No statistically significant difference (p=0.73) was observed in the mean rectal viable CT load of women with an indication for rectal testing (n=9) and without (n=15), 3.20 log10 CT/mL (range 2.06–4.36) and 3.38 log10 CT/mL (range 1.16–6.22), respectively. CT culture yielded positive test results from rectal swabs in 22.6% (12/53) of rectal CT NAAT-diagnosed women. Of women with viable rectal CT by V-PCR (n=24), 50% (12/24) were positive by CT culture.ConclusionsOverall, the detection of high rectal viable CT loads in this study indicates that rectal CT in some women might represent a currently ongoing infection rather than just the presence of remnant DNA from dead bacteria or only contamination from an active vaginal CT infection.

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Chlamydia Trachomatis Infection and Ectopic Pregnancy

Indonesian Journal of Obstetrics and Gynecology ◽

10.32771/inajog.v8i3.1136 ◽

2020 ◽

pp. 142-150

Author(s):

Septian Sima ◽

Nusratuddin Abdullah ◽

Sharvianty Arifuddin ◽

Trika Irinta ◽

Telly Tessy

Keyword(s):

Polymerase Chain Reaction ◽

Chlamydia Trachomatis ◽

Ectopic Pregnancy ◽

Cross Sectional Study ◽

Chain Reaction ◽

Cross Sectional ◽

Chlamydia Trachomatis Infection ◽

Serum Igg ◽

Polymerase Chain ◽

Ectopic Pregnancies

Objective: To determine the relationship of Chlamydia trachomatis infection in patient with ruptured ectopic pregnancy through examination of endocervical swabs, tubal tissue using Polymerase chain reaction (PCR) and Enzyme linked immunoabsorb and assay (ELISA) serum IgG antibodies Specific to Chlamydia trachomatis.Method: This study was an observational analytic study with a cross sectional study design. In this study, there were 50 participants consisting of 25 ruptured ectopic pregnancy patients and 25 non-ruptured ectopic pregnancy patients who underwent treatment at Dr.Wahidin Sudirohusodo Hospital as well as networking hospitals at the Universitas Hasanuddin in Makassar City.Results: The results showed that Chlamydia trachomatis infection in patients with ruptured ectopic pregnancies was found to be 84% positive in tubal tissue, 72% with endocervical swabs and 64% with serum examination. There was a significant relationship between chlamydial tracheal infection obtained through examination of tubal tissue, endocervical swab and specific serum IgG in patients with ruptured ectopic pregnancy (p <0.001).Conclusion: Based on the results of the study, it can be concluded that Chlamydia trachomatis infection can significantly affect the occurrence of ruptured ectopic pregnancy.Keywords: Chlamydia trachomatis, endocervical swab, ruptured ectopic pregnancy, serum IgG, tubal tissue. Abstrak Tujuan : Untuk mengetahui hubungan infeksi Chlamydia trachomatis pada pasien penderita Kehamilan Ektopik Terganggu (KET) melalui swab endoserviks dan jaringan tuba menggunakan pemeriksaan Polymerase Chain Reaction (PCR) dan Enzyme linked immunoabsorb and assay (ELISA).Metode : Penelitian ini merupakan studi analitik observasional dengan desain studi potong lintang. Pada penelitian ini terdapat 50 orang partisipan yang terdiri atas 25 orang pasien kehamilan ektopik terganggu dan 25 orang pasien non-kehamilan ektopik terganggu yang menjalani pengobatan di RSUP Dr.Wahidin Sudirohusodo serta RS jejaring Universitas Hasanuddin di Kota Makassar.Hasil : Hasil penelitian menunjukkan bahwa infeksi Chlamydia trachomatis pada pasien dengan kehamilan ektopik tergangu didapatkan sebesar 84% positif di jaringan tuba, 72% dengan swab endoserviks dan 64% dengan pemeriksaan serum. Terdapat hubungan yang signifikan antara infeksi klamidia trakomatis yang didapatkan melalui pemeriksaan jaringan tuba, swab endoserviks maupun serum IgG spesifik pasien kehamilan ektopik terganggu (p<0,001).Kesimpulan : Berdasarkan hasil penelitian, dapat disimpulkan bahwa infeksi Chlamydia trachomatis secara signifikan dapat mempengaruhi terjadinya kehamilan ektopik terganggu.Kata kunci : chlamydia trachomatis, jaringan tuba, kehamilan ektopik terganggu, swab endoserviks, serum IgG

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Unexplained Increase in Death Rate during COVID19 Pandemic Mistakenly Attributed to Malaria

European Journal of Medical and Health Sciences ◽

10.24018/ejmed.2020.2.6.545 ◽

2020 ◽

Vol 2 (6) ◽

Author(s):

Ibrahim Abdelrhim Ali ◽

Alaaeldeen Mohammed Ahmed Abdeldafia ◽

Abrar Bakry Elmalik ◽

Mohamed Eltayieb Elawad

Keyword(s):

Health Care ◽

Recovery Period ◽

Cross Sectional Study ◽

Blood Film ◽

Cross Sectional ◽

Seek Health Care ◽

Negative Results ◽

Peripheral Blood Film ◽

Males And Females ◽

Positive Results

Background: Since the emergence of COVID-19 pandemic, the world has faced many challenges. In Sudan, Gizera state has shown mysterious symptoms to the residents of a village and has been diagnosed with malaria. Aim: The study was aimed to disprove the diagnosis of malaria, and to find another more convincing explanation that fits with the general features of the disease. Method: A descriptive cross-sectional study was conducted using an electronic questionnaire designed by Google form. Result: The respondents was 402, the distribution of males and females was 51.7% and 48.3%, respectively. 83% were suffered the symptoms in May and June. The most common symptoms were headache, fever, fatigue and joint pain, back pain, sore throat and anosmia, at 60.7%, 49.8%, 47.3%, 33.3%, 37.3%, and 23.3%, respectively. 61.4% seeked health care, 99.9% performed peripheral blood film for malaria and 83% were positive. 77% of those with positive result, 72% of those with negative results, 62% of those who did not seek health care suffered similar symptoms, mainly headache. Also, anosmia is 30%, and 25% for those who have seeked and have not seeked health care, respectively. The recovery period was less than a week in 49% of those with positive results, 57% of those with negative results, and 63% of those who did not seek health care. It was more than two weeks in 9%, 12%, and 4% in the positive, negative, and uncaring, respectively. 56.5% did not notice the mosquitoes in that period, 31% and 11.7% noticed that and did not know, respectively. Conclusion: The most likely diagnosis of these mysterious symptoms is COVID-19.

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Urine Specimens from Pregnant and Nonpregnant Women Inhibitory to Amplification of Chlamydia trachomatis Nucleic Acid by PCR, Ligase Chain Reaction, and Transcription-Mediated Amplification: Identification of Urinary Substances Associated with Inhibition and Removal of Inhibitory Activity

Journal of Clinical Microbiology ◽

10.1128/jcm.36.11.3122-3126.1998 ◽

1998 ◽

Vol 36 (11) ◽

pp. 3122-3126 ◽

Cited By ~ 102

Author(s):

J. Mahony ◽

S. Chong ◽

D. Jang ◽

K. Luinstra ◽

M. Faught ◽

...

Keyword(s):

Nucleic Acid ◽

Chlamydia Trachomatis ◽

Pregnant Women ◽

False Negative ◽

Nucleic Acid Amplification ◽

Urine Specimen ◽

Chain Reaction ◽

Ligase Chain Reaction ◽

Negative Results ◽

Urine Specimens

The presence of endogenous amplification inhibitors in urine may produce false-negative results for the detection of Chlamydia trachomatis nucleic acids by tests such as PCR, ligase chain reaction (LCR), and transcription-mediated amplification (TMA). Consecutive urine specimens from 101 pregnant women and 287 nonpregnant women submitted for urinalysis were processed for C. trachomatis detection. Aliquots were spiked with the equivalent of one C. trachomatis elementary body and were tested by three commercial assays: AMPLICOR CT/NG, Chlamydia LCX, and Chlamydia TMA. The prevalence of inhibitors resulting in complete inhibition of amplification was 4.9% for PCR, 2.6% for LCR, and 7.5% for TMA. In addition, all three assays were partially inhibited by additional urine specimens. Only PCR was more often inhibited by urine from pregnant women than by urine from nonpregnant women (9.9 versus 3.1%;P = 0.011). A complete urinalysis including dipstick and a microscopic examination was performed. Logistic regression analysis revealed that the following substances were associated with amplification inhibition: beta-human chorionic gonadotropin (odds ratio [OR], 3.3) and crystals (OR, 3.3) for PCR, nitrites for LCR (OR, 14.4), and hemoglobin (OR, 3.3), nitrites (OR, 3.3), and crystals (OR, 3.3) for TMA. Aliquots of each inhibitory urine specimen were stored at 4 and −70°C overnight or were extracted with phenol-chloroform and then retested at dilutions of 1:1, 1:4, and 1:10. Most inhibition was removed by storage overnight at 4 or −70°C and a dilution of 1:10 (84% for PCR, 100% for LCR, and 92% for TMA). Five urine specimens (three for PCR and two for TMA) required phenol-chloroform extraction to remove inhibitors. The results indicate that the prevalence of nucleic acid amplification inhibitors in female urine is different for each technology, that this prevalence may be predicted by the presence of urinary factors, and that storage and dilution remove most of the inhibitors.

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Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

Anti-Infective Agents ◽

10.2174/2211352518999200629165108 ◽

2020 ◽

Vol 18 ◽

Author(s):

Pegah Shakib ◽

Mohammad Reza Zolfaghari

Keyword(s):

Streptococcus Pneumoniae ◽

Real Time ◽

Real Time Pcr ◽

False Negative ◽

Cross Sectional Study ◽

Laboratory Culture ◽

Cross Sectional ◽

Negative Results ◽

False Negative Results ◽

Pcr Method

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

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Prevalence of urogenital, anal, and pharyngeal infections with Chlamydia trachomatis, Neisseria gonorrhoeae, and Mycoplasma genitalium: a cross-sectional study in Reunion island

BMC Infectious Diseases ◽

10.1186/s12879-021-05801-9 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

A. Calas ◽

N. Zemali ◽

G. Camuset ◽

J. Jaubert ◽

R. Manaquin ◽

...

Keyword(s):

Chlamydia Trachomatis ◽

Neisseria Gonorrhoeae ◽

Mycoplasma Genitalium ◽

Cross Sectional Study ◽

Sectional Study ◽

Transmitted Infection ◽

Cross Sectional ◽

Sexually Transmitted ◽

Urogenital Infection ◽

High Prevalence

Abstract Background Recommendations for sexually transmitted infection (STI) screening vary significantly across countries. This study evaluated the prevalence of urogenital and extragenital infections with Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG), and Mycoplasma genitalium (MG) in patients visiting a French STI clinic in the Indian Ocean region to determine whether current STI screening practices should be updated. Methods This cross-sectional study examined all patients who visited the STI clinic between 2014 and 2015. Triplex polymerase chain reaction screening for CT, NG, and MG was performed on urine, vaginal, pharyngeal, and anal specimens (FTD Urethritis Basic Kit, Fast Track Diagnostics, Luxembourg). Results Of the 851 patients enrolled in the study, 367 were women (367/851, 43.2%) and 484 were men (484/851, 56.0%). Overall, 826 urogenital specimens (826/851, 97.1%), 606 pharyngeal specimens (606/851, 71.2%), and 127 anal specimens (127/851, 14.9%) were taken from enrolled patients. The prevalence of urogenital CT and MG was high in women ≤25 years (19/186, 10.21%; 5/186, 2.69%) and in men who have sex with women ≤30 years (16/212, 7.54%; 5/212, 2.36%). Among patients with urogenital CT infection, 13.7% (7/51) had urethritis. All patients with urogenital MG infection were asymptomatic. Men who have sex with men had a high prevalence of pharyngeal CT (2/45, 4.44%) and NG (3/44, 6.81%) and a high prevalence of anal CT (2/27, 7.41%), NG (2/27, 7.40%), and MG (1/27, 3.70%). After excluding patients with concomitant urogenital infection, extragenital infections with at least 1 of the 3 pathogens were found in 20 swabs (20/91, 21.9%) taken from 16 patients (16/81, 19.7%), all of them asymptomatic. Conclusions Routine multisite screening for CT, NG, and MG should be performed to mitigate the transmission of STIs in high-risk sexually active populations.

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Evaluation of SARS-CoV-2 antigen-based rapid diagnostic kits in Pakistan: formulation of COVID-19 national testing strategy

Virology Journal ◽

10.1186/s12985-021-01505-3 ◽

2021 ◽

Vol 18 (1) ◽

Author(s):

Umar Saeed ◽

Sara Rizwan Uppal ◽

Zahra Zahid Piracha ◽

Azhar Rasheed ◽

Zubair Aftab ◽

...

Keyword(s):

Gold Standard ◽

Injection Drug Users ◽

Drug Users ◽

Diagnostic Testing ◽

False Negative ◽

Cross Sectional Study ◽

Nasopharyngeal Swab ◽

Rt Pcr ◽

Cross Sectional ◽

Negative Results

AbstractRapid diagnosis of SARS-CoV-2 during pandemic enables timely treatment and prevention of COVID-19. Evaluating the accuracy and reliability of rapid diagnostic testing kits is crucial for surveillance and diagnosis of SARS-CoV-2 infections in general population, injection drug users, multi-transfused populations, healthcare workers, prisoners, barbers and other high risk populations. The aim of this study was to evaluate performance and effectiveness of nasopharyngeal swab (NSP) and saliva based rapid antigen detection testing kits in comparison with USFDA approved triple target gold standard real-time polymerase chain reaction. A cross-sectional study was conducted on 33,000 COVID-19 suspected patients. From RT-PCR positive patients, nasopharyngeal swab (NSP) and saliva samples were obtained for evaluation of rapid COVID-19 testing kits (RDT). 100/33,000 (0.3%) of specimens were RT-PCR positive for SARS-CoV-2. Among RT-PCR positive, 62% were males, 34% were females, and 4% were children. The NSP-RDT (Lepu Medical China) analysis revealed 53% reactivity among males, 58% reactivity among females, and 25% reactivity among children. However saliva based RDT (Lepu Medical China) analysis showed 21% reactivity among males and 23% among females, and no reactivity in children. False negative results were significantly more pronounced in saliva based RDT as compared to NSP-RDT. The sensitivity of these NSP-RDT and saliva based RDT were 52% and 21% respectively. The RDTs evaluated in this study showed limited sensitivities in comparison to gold standard RT-PCR, indicating that there is a dire need in Pakistan for development of suitable testing to improve accurate COVID-19 diagnosis in line with national demands.

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A Retrospective Cross-Sectional Study of the Utility of Cartridge-Based Nucleic Acid Amplification Test in Diagnosis of Pulmonary and Extrapulmonary Tuberculosis in People Living with HIV in Second Highest HIV Prevalent State in India

International Journal of Recent Surgical and Medical Sciences ◽

10.1055/s-0041-1730242 ◽

2021 ◽

Author(s):

Tade Bagbi ◽

Ningthoukhongjam Reema ◽

S. Bhagyabati Devi ◽

Thangjam Gautam Singh ◽

Mohammad Jaleel ◽

...

Keyword(s):

Nucleic Acid ◽

Cross Sectional Study ◽

Nucleic Acid Amplification Test ◽

Nucleic Acid Amplification ◽

Rifampicin Resistance ◽

Sputum Smear ◽

Smear Microscopy ◽

Sectional Study ◽

Cross Sectional ◽

Smear Negative

Abstract Introduction Tuberculosis (TB) in people living with human immunodeficiency virus (PLHIV) is difficult to diagnose due to fewer organisms in sputum and extrapulmonary samples. Sputum culture takes 4 to 8 weeks for growth of the mycobacteria. Delayed treatment for TB in PLHIV leads to increased mortality. This study evaluated cartridge-based nucleic acid amplification test (CBNAAT) as a diagnostic tool for diagnosis of pulmonary TB (PTB) and extrapulmonary TB (EPTB) in PLHIV in the second most HIV prevalent state in India and for comparing its efficacy between Ziehl–Neelsen (ZN) staining sputum smear–positive and sputum smear–negative TB. Methods This cross-sectional study was conducted in RIMS, Imphal, with 167 PLHIV patients, age 15 years or older, having signs and symptoms of TB. Appropriate samples for sputum microscopy and CBNAAT were sent. Conclusion The overall sensitivity of sputum smear for acid-fast bacillus (AFB) was found to be 30.71% and that of CBNAAT was 38.57%. Sensitivity of CBNAAT for sputum smear–positive and sputum smear–negative TB was 100 and 11.3%, respectively. Sensitivity of ZN smear for AFB of EPTB sample was 48.1% and that of CBNAAT was 59.25%. In both PTB and EPTB, CBNAAT showed an increase in diagnosis of microbiologically confirmed PTB cases by 7.8 and 11.1%, respectively, over and above the cases diagnosed by ZN smear microscopy. Rifampicin resistance was detected in five patients. We conclude that CBNAAT is a rapid test with better sensitivity in diagnosis of PTB and EPTB in PLHIV, compared with ZN smear microscopy. It detects rifampicin resistance for multidrug-resistant TB and helps in early treatment intervention.

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