Mixed Infection Rate Research Articles

Development of a Multiplex Quantitative PCR for Detecting Porcine Epidemic Diarrhea Virus, Transmissible Gastroenteritis Virus, and Porcine Deltacoronavirus Simultaneously in China.

Porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and porcine deltacoronavirus (PDCoV) belong to the category of swine enteric coronavirus that cause acute diarrhea in piglets, which has resulted in massive losses to the pig husbandry. Therefore, a sensitive and rapid detection method which can differentially detect these viruses that lead to mixed infections in clinical cases, is urgently needed. According to the conserved regions of the PEDV M gene, TGEV S gene, and PDCoV N gene, and the reference gene of porcine (β-Actin), we designed new specific primers and probes for the multiplex qPCR assay capable of simultaneously detecting three RNA viruses. This method, with a great specificity, did not cross-react with the common porcine virus. Moreover, the limit of detection of the method we developed could reach 10 copies/μL ,and the intra- and inter-group coefficients of variation of it below 3%. Applying this assay to detect 462 clinical samples which were collected in 2022-2023, indicated that the discrete positive rates of PEDV, TGEV, and PDCoV were 19.70%, 0.87%, and 10.17%, respectively. The mixed infection rates of PEDV/TGEV, PEDV/PDCoV, TGEV/PDCoV, and PEDV/TGEV/PDCoV were 3.25%, 23.16%, 0.22%, and 11.90%, respectively. All in all, the multiplex qPCR assay we developed as a tool for differential and rapid diagnosing can be put on the active prevention and control of PEDV, TGEV, and PDCoV, , which can create great value in the diagnosis of swine diarrhea diseases.

Clinical application value of metagenomic second-generation sequencing technology in hematologic diseases with and without transplantation.

Hematological patients are at risk of infections. It is unknown whether the pathogenic microbial spectrum differs between HSCT and non-HSCT patients, and whether metagenomic next-generation sequencing (mNGS) of peripheral blood can be used as a substitute test specimen such as alveolar lavage. A retrospective study was conducted to evaluate the clinical application value of mNGS in hematological patients with and without HSCT. Viruses were prevalent pathogens in both non-HSCT (44%) and HSCT (45%) patients, chiefly human cytomegalovirus and Epstein-Barr virus. In non-HSCT patients, Gram-negative bacilli accounted for 33% (predominantly Klebsiella pneumonia), and Gram-positive cocci accounted for 7% (predominantly Enterococcus faecium) of pathogens. However, in HSCT patients, Gram-negative bacilli accounted for 13% (predominantly Stenotrophomonas maltophilia), and Gram-positive cocci accounted for 24% (predominantly Streptococcus pneumonia) of pathogens. Mucor was the most common fungu s in two groups. The positive rate of pathogens by mNGS was 85.82%, higher than conventional detection (20.47%, P < 0.05). Mixed infection accounted for 67.00%, among which the mixed infection of bacteria and virus (25.99%) was the most common. 78 cases had pulmonary infection, the positive rate of traditional laboratory tests was 42.31% (33/78), and of mNGS in peripheral blood was 73.08% (57/78), showing a statistical difference (P = 0.000). The non-HSCT patients had a higher frequency of Klebsiella pneumonia (OR=0.777, 95% CI, 0.697-0.866, P = 0.01) and Torque teno virus (OR=0.883, 95% CI, 0.820-0.950, P = 0.031) infections than HSCT patients, while the rates of Streptococcus pneumonia (OR=12.828, 95% CI, 1.378-119.367, P = 0.016), Candida pseudosmooth (OR=1.100, 95% CI, 0.987-1.225, P = 0.016), human betaherpesvirus 6B (OR=6.345, 95% CI, 1.105-36.437, P = 0.039) and human polyomavirus 1 (OR=1.100, 95% CI, 0.987-1.225, P = 0.016) infections were lower. Leishmania could be detected by mNGS. mNGS of peripheral blood can be used as a substitute test method for hematological patients with pulmonary infection, the detection rate of mixed infections by mNGS was high, and mNGS has high clinical recognition rate and sensitivity in pathogen detection, and provides a basis for guiding the anti-infective treatment in hematological diseases with symptoms such as fever.

Whole-Genome Sequencing Exhibits Better Diagnostic Performance than Variable-Number Tandem Repeats for Identifying Mixed Infections of Mycobacterium tuberculosis.

Mixed infections of Mycobacterium tuberculosis, defined as the coexistence of multiple genetically distinct strains within a single host, have been associated with unfavorable treatment outcomes. Different methods have been used to detect mixed infections, but their performances have not been carefully evaluated. To compare the sensitivity of whole-genome sequencing (WGS) and variable-number tandem repeats (VNTR) typing to detect mixed infections, we prepared 10 artificial samples composed of DNA mixtures from two strains in different proportions and retrospectively collected 1,084 clinical isolates. The limit of detection (LOD) for the presence of a minor strain was 5% for both WGS and VNTR typing. The overall clinical detection rate of mixed infections was 3.7% (40/1,084) for the two methods combined, WGS identified 37/1,084 (3.4%), and VNTR typing identified 14/1,084 (1.3%), including 11 also identified by WGS. Multivariate analysis demonstrated that retreatment patients had a 2.7 times (95% confidence interval [CI], 1.2 to 6.0) higher risk of mixed infections than new cases. Collectively, WGS is a more reliable tool to identify mixed infections than VNTR typing, and mixed infections are more common in retreated patients. IMPORTANCE Mixed infections of M. tuberculosis have the potential to render treatment regimens ineffective and affect the transmission dynamics of the disease. VNTR typing, currently the most widely used method for the detection of mixed infections, detects mixed infections only by interrogating a small fraction of the M. tuberculosis genome, which necessarily limits sensitivity. With the introduction of WGS, it became possible to study the entire genome, but no quantitative comparison has yet been undertaken. Our systematic comparison of the ability of WGS and VNTR typing to detect mixed infections, using both artificial samples and clinical isolates, revealed the superior performance of WGS at a high sequencing depth (~100×) and found that mixed infections are more common in patients being retreated for tuberculosis (TB) in the populations studied. This provides valuable information for the application of WGS in the detection of mixed infections and the implications of mixed infections for tuberculosis control.

Malaria prevalence in Pakistan: A systematic review and meta-analysis (2006–2021)

Malaria is one of the major public health issues globally. Malaria infection spreads through mosquito bites from infected female Anopheles mosquitoes. This study aims to conduct a systematic review and meta-analysis on malaria prevalence in Pakistan from 2006 to 2021. We searched PubMed, Science Direct, EMBASE, EMCare, and Google Scholar to acquire data on the prevalence of malaria infections. We performed a meta-analysis with a random-effects model to obtain the pooled prevalence of malaria, Plasmodium vivax, and Plasmodium falciparum. Meta-analysis was computed using R 4.1.2 Version statistical software. I2 and time series analysis were performed to identify a possible source of heterogeneity across studies. A funnel plot and the Freeman-Tukey Double Arcsine Transformed Proportion were used to evaluate the presence of publication bias. Out of the 315 studies collected, only 45 full-text articles were screened and included in the final measurable meta-analysis. Pooled malaria prevalence in Pakistan was 23.3%, with Plasmodium vivax, Plasmodium falciparum, and mixed infection rates of 79.13%, 16.29%, and 3.98%, respectively. Similarly, the analysis revealed that the maximum malaria prevalence was 99.79% in Karachi and the minimum was 1.68% in the Larkana district. Amazingly, this systematic review and meta-analysis detected a wide variation in malaria prevalence in Pakistan. Pakistan's public health department and other competent authorities should pay close attention to the large decrease in mosquito populations to curb the infection rate.

Development of a TaqMan-Probe-Based Multiplex Real-Time PCR for the Simultaneous Detection of African Swine Fever Virus, Porcine Circovirus 2, and Pseudorabies Virus in East China from 2020 to 2022

African swine fever virus (ASFV), porcine circovirus 2 (PCV2), and pseudorabies virus (PRV) are important DNA viruses that cause reproductive disorders in sows, which result in huge losses in pig husbandry, especially in China. The multiplex qPCR assay could be utilized as a simultaneous diagnostic tool for field-based surveillance and the control of ASFV, PCV2, and PRV. Based on the conserved regions on the p72 gene of ASFV, the Cap gene of PCV2, the gE gene of PRV, and the porcine endogenous β-Actin gene, the appropriate primers and probes for a multiplex TaqMan real-time PCR test effective at concurrently detecting three DNA viruses were developed. The approach demonstrated high specificity and no cross-reactivity with major pathogens related to swine reproductive diseases. In addition, its sensitivity was great, with a detection limit of 101 copies/L of each pathogen, and its repeatability was excellent, with intra- and inter-group variability coefficients of <2%. Applying this assay to detect 383 field specimens collected from 2020 to 2022, the survey data displayed that the ASFV, PCV2, and PRV single infection rates were 22.45%, 28.46%, and 2.87%, respectively. The mixed infection rates of ASFV + PCV2, ASFV + PRV, PCV2 + PRV, and ASFV + PCV2 + PRV were 5.22%, 0.26%, 1.83%, and 0.26%, respectively. Overall, the assay established in this study provides an effective tool for quickly distinguishing the viruses causing sow reproductive disorders, suggesting its huge clinical application value in the diagnosis of swine diseases.

Epidemiology of infection with lung worms in sheep in Tikrit city, Salah Al Deen province

1.The study area spend Tikrit in Salah Aden province, and followed him from the aspects and villages and tubing such as Al-Alam district , Hawi Albu-Ajeel, Al-Naama areas, a series Hamrin mountains east of the Tigris River, stretching from north to south and Al Awja district, Aouinat, Mahzm, Al-Hamra and al-Jazeera to Lake Tharthar west of the Tigris River.&#x0D; &#x0D; were collected 1298 fecal sample from sheep and goats, of which 734 samples of sheep and 564 samples of the goats as "the beginning of the month of January until the end of the first December for the year (2014), have been examined and these samples to look for larvae of the lung worms in sheep of five races are Dictyocaulus, Muellerius, Protostrongylus, Cystocaulus and Neostrongylus test result showed the following:&#x0D; &#x0D; A.The effect of infection for the seasons of the year. The highest proportion of infection in the early winter (December first) while the lowest rate of infection in the mid-summer (July).&#x0D; &#x0D; Ratio of infection for genus lung worms. The rate of infection genus Dictyocaulus 21.5%, the highest rate while the rate of infection genus Neostrongylus 2.1%, representing the lowest percentage of infection among the five races the rest of the races where concatenated infection rates downward Muellerius 13.6% and then Protostrongylus 10.9 % then Cystocaulus .&#x0D; Ratio of infection single and mixed. Single scored infection rate of 13.79% and mixed infection rate of 15.48% during the year (2014).&#x0D; Show that the proportion of infection to the search area along the Tigris River is higher than the rest of the other search areas due to contamination of pasture and increase the availability of moisture and Alamadaiv moderation in all seasons of the year.&#x0D;

Comparison of the clinical features of human bocavirus and metapneumovirus lower respiratory tract infections in hospitalized children in Suzhou, China.

We compared the clinical data of hospitalized children with lower respiratory tract infections caused by human bocavirus (HBoV) and human metapneumovirus (hMPV). In total, 8,430 children admitted to the Department of Respiration, Children's Hospital of Soochow University for lower respiratory tract infections from January 2017 to October 2021 were enrolled. Seven common respiratory viruses, including respiratory syncytial virus, influenza virus A, influenza virus B, parainfluenza virus (PIV) I, PIV II, PIV III, and adenovirus, were detected by direct immunofluorescence assay, whereas human rhinovirus and hMPV were detected by reverse transcription-polymerase chain reaction. Mycoplasma pneumoniae (MP) and HBoV were detected by real-time fluorescence quantitative polymerase chain reaction. Bacteria was detected in blood, nasopharyngeal secretion, bronchoalveolar lavage specimen or pleural fluid by culture. In parallel, MP was detected by enzyme-linked immunosorbent assay. In addition, we performed metagenomic testing of alveolar lavage fluid from some of the patients in our study. The detection rate of HBoV was 6.62% (558/8430), whereas that of hMPV was 2.24% (189/ 8430). The detection rate of HBoV was significantly higher in children aged 1 to <3 years than in other age groups, but there were no significant differences in positivity rates for hMPV by age. Before 2020, the incidence of HBoV infection peaked in summer and autumn, whereas that of hMPV peaked in spring. The epidemiology of both HBoV and hMPV has changed because of the impact of the novel coronavirus. Among the positive cases, the HBoV mixed infection rate was 51.6%, which was similar to that for hMPV mixed infection (44.4%). Comparing clinical characteristics between HBoV and hMPV single infection, the median age of children was 17 months in the HBoV group and 11 months in the hMPV group. In the HBoV single infection group, 31 patients (11.5%) had pulse oxygen saturation of less than 92% on admission, 47 (17.4%) had shortness of breath, and 26 (9.6%) presented with dyspnea. Meanwhile, four patients (3.8%) in the hMPV single infection group had pulse oxygen saturation of less than 92% on admission, eight (7.6%) displayed shortness of breath, and three (2.9%) had dyspnea. The proportion of patients requiring mechanical ventilation and the rate of PICU admission were higher in the HBoV group than in the hMPV group. The prevalence of HBoV infection is higher than that of hMPV infection in children with lower respiratory tract infection in Suzhou, and HBoV is more likely to cause severe infection than hMPV. Public health interventions for COVID-19 outbreaks have affected the prevalence of HBoV and hMPV.

Development of a TaqMan-Probe-Based Multiplex Real-Time PCR for the Simultaneous Detection of Porcine Circovirus 2, 3, and 4 in East China from 2020 to 2022.

Porcine circovirus disease (PCVD) caused by porcine circovirus (PCV) is an important swine disease that is characterized by porcine dermatitis, nephropathy syndrome, and reproductive disorders in sows. However, disease caused by PCV2, PCV3, or PCV4 is hard to distinguish, so a rapid and sensitive detection method is urgently needed to differentiate these three types. In this study, four pairs of specific primers and the corresponding probes for PCV 2, -3, and -4, and porcine endogenous gene β-Actin as the positive internal reference index, were designed to establish a TaqMan multiplex real-time PCR (qPCR) assay for the simultaneous differential diagnosis of different types of viruses. The results showed that this assay has good specificity and no cross-reactivity with other important porcine viral pathogens. Furthermore, it has high sensitivity, with a detection limit of 101 copies/μL, and good reproducibility, with intra- and inter-group coefficients of variation below 2%. Subsequently, 535 clinical samples of suspected sow reproductive disorders collected from Shandong, Zhejiang, Anhui, and Jiangsu provinces from 2020 to 2022 were analyzed using the established assay. The results showed that the individual positive rates of PCV2, PCV3, and PCV4 were 31.03%, 30.09%, and 30.84%, respectively; the mixed infection rates of PCV2 and PCV3, PCV2 and PCV4, and PCV3 and PCV4 were 31.03%, 30.09%, and 30.84%, respectively; the mixed infection rate of PCV2, PCV3, and PCV4 was 28.22%. This indicated that this assay provides a convenient tool for the rapid detection and differentiation of PCV2, PCV3, and PCV4 in pig farms in East China. Our findings highlight that there are different types of porcine circovirus infection in pig farms in East China, which makes pig disease prevention and control difficult.

Analysis of non-bacterial respiratory pathogens in children in Ningbo City from 2019 to 2021

Objective: To investigate the distribution characteristics of respiratory non-bacterial pathogens in children in Ningbo from 2019 to 2021. Methods: A retrospective analysis was performed on 23 733 children with respiratory tract infection who visited the department of pediatrics of Ningbo Women and Children's Hospital from July 2019 to December 2021. There were 13 509 males (56.92%) and 10 224 females (43.08%), with an age range of 1 day to 18 years old. There were 981 cases in the neonatal group (younger than 1 month old), 5 880 cases in the infant group (1 month to younger than 1 year old), 6 552 cases in the toddler group (1 to younger than 3 years old), 7 638 cases in the preschool group (3 to younger than 7 years old), and 2 682 cases in the school-age group (7 to 18 years old). Thirteen respiratory pathogens were detected by multiple polymerase chain reaction (PCR) based on capillary electrophoresis, and SPSS 23.0 software was used for statistical analysis of the results, the count data were expressed as percentages, and the χ2 test was used for comparison between groups. Results: Of the 23 733 specimens, 13 330 were positive for respiratory pathogens, with a total positive rate of 56.17%. The positive rates of human rhinovirus (HRV) 24.05% (5 707/23 733), human respiratory syncytial virus (HRSV) 10.45% (2 480/2 3733) and mycoplasma pneumoniae (Mp) 7.03% (1 668/23 733) were in the first three. The positive rates of pathogens in the male and female children were 57.47% (7 763/13 509) and 54.45% (5 567/10 224), respectively, and the difference was statistically significant (χ2=21.488, P<0.001). The positive rates in the neonatal group, infant group, toddler group, preschool group, and school-age group were 31.80% (312/981), 54.71% (3 217/5 880), 63.23% (4 143/6 552), 59.83% (4 570/7 638), 40.57% (1 088/2 682), respectively, and the difference among the groups was statistically significant (χ2=681.225, P<0.001). The single infection rate was 47.43% (11 256/23 733), the mixed infection rate of two or more pathogens was 8.74% (2 074/23 733), most of which were mixed infections of two pathogens. HRV, HADV, HCOV, Ch disseminated in the whole year. HRSV, HMPV, Boca, HPIV occurred mostly in fall and winter. The positive rates of FluA, FluB, Mp were at a low level after the corona virus disease 2019 (COVID-19) epidemic (2020 and 2021). The positive rates of FluA, H1N1, H3N2, FluB, HADV, Mp in 2020 were significantly lower than in 2019 (P<0.05). The positive rates of HPIV, HRV, HCOV, Ch in 2020 were significantly higher than in 2019 (P<0.05). The positive rates of FluA, H1N1, H3N2, HPIV, HCOV, Mp, Ch in 2021 were significantly lower than in 2020 (P<0.05). The positive rates of Boca, HMPV, HRSV in 2021 were significantly higher than in 2020 (P<0.05). Conclusion: From 2019 to 2021, the main non-bacterial respiratory pathogens of children in Ningbo City were Mp and HRV, and the detection rates of respiratory pathogens varied among different ages, seasons and genders.

The evaluation of next-generation sequencing assisted pathogenic detection in immunocompromised hosts with pulmonary infection: A retrospective study.

Pulmonary infections are frequent in immunocompromised hosts (ICH), and microbial detection is difficult. As a new method, next-generation sequencing (NGS) may offer a solution. This study aimed to assess the impact of NGS-assisted pathogenic detection on the diagnosis, treatment, and outcomes of ICH complicated by pulmonary infection and radiographic evidence of bilateral diffuse lesions. This study enrolled 356 patients with ICH complicated by pulmonary infection that were admitted to Zhongshan Hospital, Fudan University, from November 17, 2017, to November 23, 2018, including 102 and 254 in the NGS and non-NGS groups, respectively. Clinical characteristics, detection time, rough positive rate, effective positive rate, impact on anti-infective treatment plan, 30-day/60-day mortality, and in-hospital mortality were compared. NGS-assisted pathogenic detection reduced detection time (28.2 h [interquartile range (IQR) 25.9-29.83 h] vs. 50.50 h [IQR 47.90-90.91 h], P < 0.001), increased positive rate, rate of mixed infection detected, effective positive rate, and proportion of antibiotic treatment modification (45.28% vs. 89.22%, 4.72% vs. 51.96%, 21.65% vs. 64.71%, 16.54% vs. 46.08%, P < 0.001). The NGS group had a significantly lower 60-day mortality rate (18.63% vs. 33.07%, P = 0.007). The difference in the Kaplan-Meier survival curve was significant (P = 0.029). After multivariate logistic regression, NGS-assisted pathogenic detection remained a significant predictor of survival (OR 0.189, confidence interval [CI], 0.068-0.526). NGS-assisted pathogenic detection may improve detection efficiency and is associated with better clinical outcomes in these patients.

Development and application of a multiplex qPCR assay for the detection of duck circovirus, duck Tembusu virus, Muscovy duck reovirus, and new duck reovirus.

A multiplex qPCR assay was developed to simultaneously detect duck circovirus (DuCV), duck Tembusu virus (DTMUV), Muscovy duck reovirus (MDRV), and novel duck reovirus (NDRV), but it did not amplify other viruses, including duck virus enteritis (DVE), infectious bursal disease virus (IBDV), avian reovirus (ARV), H5 avian influenza virus (H5 AIV), H7 avian influenza virus (H7 AIV), H9 avian influenza virus (H9 AIV), Newcastle disease virus (NDV), and Muscovy duck parvovirus (MDPV), and the detection limit for DuCV, DTMUV, MDRV, and NDRV was 1.51 × 101 copies/μL. The intra- and interassay coefficients of variation were less than 1.54% in the repeatability test with standard plasmid concentrations of 1.51 × 107, 1.51 × 105, and 1.51 × 103 copies/μL. The developed multiple qPCR assay was used to examine 404 clinical samples to verify its practicability. The positivity rates for DuCV, DTMUV, MDRV, and NDRV were 26.0%, 9.9%, 4.0%, and 4.7%, respectively, and the mixed infection rates for DuCV + DTMUV, DuCV + MDRV, DuCV + NDRV, MDRV + NDRV, DTMUV + MDRV, and DTMUV + NDRV were 2.7%, 1.2%, 1.2%, 1.0%, 0.5%, and 0.7%, respectively.

Human rotavirus VP4 and VP7 genetic diversity and detection of GII norovirus in Ibadan as Nigeria introduces rotavirus vaccine.

ObjectiveThis cross-sectional study investigated the circulating strains of rotavirus and screened for noravirus in Ibadan, Nigeria as the country introduces the rotavirus vaccine into its national immunization program.MethodsSixty-five stool samples were collected from children younger than 5 years with clinically diagnosed diarrhea and screened for the presence of rotavirus and norovirus using RT-PCR. Rotavirus-positive samples were further analyzed to determine the G and P genotypes using semi-nested multiplex PCR.ResultsThe rates of rotavirus and norovirus positivity were 30.8% and 10.8%, respectively, whereas the rate of rotavirus and norovirus mixed infection was 4.6%. G1 was the predominant VP7 genotype, followed by G2, G9, and G1G2G9, whereas the predominant VP4 genotype was P[4], followed by P[6], P[8], and P[9]. The mixed P types P[4]P[8] and P[4]P[6] were also detected. G1P[4] was the most common VP4 and VP7 combination, followed by G2P[4], G1[P6], G1P[8], G2P[6], G2P[9], G9P[6], G2G9P[4], G2P[4]P[6], G1P[4]P[8], G2G9P[8], G1G2G9P[8], and G1[non-typable] P[non-typable], which were detected in at least 5% of the samples. Four samples had a combination of non-typable G and P types.ConclusionsIt is essential to monitor the circulation of virus strains prior to and during the implementation of the immunization program.

Patterns of the Occurrence of TYLCV and ToCV with Whitefly on Summer-Cultivated Tomato in Greenhouse in Gwangju, Gyeonggi Province

Patterns of occurrence of tomato yellow leaf curl virus (TYLCV) and tomato chlorosis virus (ToCV) with whitefly on summer-cultivated tomato in Gwangju-si, Gyeonggi Province were surveyed using multiplex reverse transcription-polymerase chain reaction in 2020. In addition, distribution of the whiteflies species and their viral transmission rates were investigated throughout the tomato growing season. The infection rates of TYLCV and ToCV increased sharply during harvest, and the single infection rates were 30.9% and 5.0%, respectively, with a mixed infection rate of the two viruses being the highest at 52.2%. Single infection with TYLCV and double infections with TYLCV and ToCV accounted for the majority with 83.1%. Bemisia tabaci were dominant over Trialeurodes vaporariorum in greenhouse grown plants, and all of the investigated B. tabaci biotypes were identified as Mediterranean (MED, formerly known as Q biotype). The transmission rate of TYLCV, detected in every sampled B. tabaci MED population, was 21.4%, and the mixed transmission rate with ToCV was 35.5%. Viruliferous MED whiteflies with ToCV showed a higher rate than that of T. vaporariorum. In the transplant stage, viruliferous rate of both TYLCV and ToCV of B. tabaci was 42.7%; this rate was highest in the harvest stage. In examination of tomato yield, the increase in the mixed infection rate of TYLCV and ToCV led to complete yield loss. When the mixed infection rate increased by 10%, the yield decreased by 405.4 kg/10a.

High infection rate of tick-borne protozoan and rickettsial pathogens of cattle in Malawi and the development of a multiplex PCR for Babesia and Theileria species identification

Malawi has an estimated cattle population of 1,884,803 heads, the indigenous Malawi zebu breed accounts for 91.2%, while the exotic and crossbred accounts for the remaining 8.8%. Although ticks and tick-borne diseases are widespread in Malawi, no molecular study has been conducted to investigate the tick-borne Anaplasmataceae and piroplasms infecting cattle. To provide an insight into the current status of tick-borne pathogens (TBPs) of cattle, a molecular survey was conducted in the central and southern regions of Malawi. A total of 191 cattle of which 132 were Malawi zebu, 44 were Holstein Friesian and 15 were Holstein-Friesian/ Malawi zebu crosses were screened for Anaplasmataceae and piroplasms using the heat shock protein groEL gene and 18S rDNA, respectively. A new 18S rDNA multiplex PCR assay was designed for Babesia and Theileria species identification without sequencing. Overall, 92.3% (n = 177) of the examined animals were infected with at least one TBP. Anaplasmataceae-positive rate was 57.6% (n = 110) while for piroplasms it was 80.1% (n = 153). The detected Anaplasmataceae were Anaplasma bovis 2.6% (n = 5), Anaplasma marginale 24.6% (n = 47), Anaplasma platys-like 13.6% (n = 26), uncharacterized Anaplasma sp. 14.1% (n = 27), and uncharacterized Ehrlichia sp. 16.2% (n = 31). The detected piroplasms were Babesia bigemina 2.6% (n = 5), Theileria mutans 73.8% (n = 141), Theileria parva 33.0% (n = 63), Theileria taurotragi 12.6% (n = 24), and Theileria velifera 53.4% (n = 102). Mixed infection rate was found in 79.6% (n = 152) of the samples analyzed. This study has shown a high burden of TBPs among cattle in Malawi which highlights the need to conceive new methods to control ticks and TBPs in order to improve animal health and productivity. The newly developed multiplex PCR assay would be a useful tool especially in resource limited settings where sequencing is not available and when mixed infections are expected.

A study on vaginitis among pregnant and non-pregnant females in Alexandria, Egypt: An unexpected high rate of mixed vaginal infection.

BackgroundMany infectious and noninfectious triggers lead to inflammation of the vagina.AimWe investigated the prevalence of causative vaginitis microorganisms in 516 pregnant and nonpregnant female volunteers. Vaginal samples were examined microscopically, cultured and tested for different pathogens.ResultsOf the participants, 310 (60.1%) were pregnant, whereas 206 (39.9%) were nonpregnant. Using Amsel's criteria and Nugent's scores, bacterial vaginosis (BV) was diagnosed in 59.1%, and the prevalence of vulvovaginal candidiasis (VVC) was 50.2% in the population. Candida infections were significantly higher in nonpregnant females (p value ≤ 0.01), and 24% of females had mixed infections. The most common mixed infection was BV and Candida spp., detected in 21% of the cases.ConclusionsBacterial vaginosis is the most common cause of vaginitis. We observed that 24% of females experienced mixed infections, and Candida albicans was the most common fungal species causing VVC. Trichomonas vaginalis prevalence was underestimated using wet mounts.

Effect of Mixed Infections with Mycobacterium tuberculosis and Nontuberculous Mycobacteria on Diagnosis of Multidrug-Resistant Tuberculosis: A Retrospective Multicentre Study in China.

BackgroundCorrect species identification is essential before initiation of TB treatment, due to substantial drug susceptibility profile differences among mycobacterial species. Given that nontuberculous mycobacteria (NTM) are frequently resistant to first-line anti-tuberculosis drugs, cases with mixed infections with Mycobacterium tuberculosis (MTB) and NTM tend to be diagnosed as multidrug-resistant tuberculosis (MDR-TB) cases. Here we report results of a retrospective multicentre study that was conducted to determine the prevalence of TB-NTM infections in previously diagnosed laboratory-confirmed multidrug-resistant tuberculosis (MDR-TB) patients using phenotypic drug susceptibility testing. The results were then used to identify risk factors associated with susceptibility to mixed infections.MethodsFrom January 2019 through December 2019, we retrospectively collected MDR-TB isolates from three TB specialised hospitals. Species identifications of isolates were performed using the MeltPro Myco assay.ResultsA total of 837 MDR-TB isolates were analysed, of which 22 isolates (2.6%) were found to contain a mixture of NTM and MTB organisms. Significant differences in prevalence rates of mixed infections across regions were observed, with prevalence rates ranging from 0.0% (0/213) in Beijing to 3.4% (12/353) in Fuzhou to 3.7% (10/271) in Guangzhou. Among the 22 patients with NTM-TB mixed infections, a total of five different mycobacterial species were identified, of which the most prevalent species was Mycobacterium intracellulare. Notably, a history of previous TB episodes correlated with higher mixed infection risk.ConclusionThe results reported here demonstrated that mixed infections with MTB and NTM occurred in approximately 3% of suspected MDR-TB patients in China. These findings raise concerns about the accuracy of molecular diagnostics-based species identification tests and draw attention to the possibility that NTM-MTB mixed infections will be misdiagnosed as MDR-TB in high TB burden settings.

Epidemiology of echinococcosis in Serthar County of Sichuan Province in 2019

To investigate the epidemiology of echinococcosis in Serthar County, Sichuan Province, so as to provide insights into optimization of echinococcosis control measures. The prevalence of human echinococcosis was surveyed among permanent residents living in Serthar County at ages of over 2 years from 2016 to 2019, and the prevalence of Echinococcus infections was surveyed in owned dogs, yaks and rodents in 2019. The epidemiological features of echinococcosis in humans and animals were descriptively analyzed. The overall prevalence of human echinococcosis was 3.72% (1 613/43 362) in Serthar County from 2016 to 2019, and the prevalence rates of cystic echinococcosis, alveolar echinococcosis and mixed infections of cystic echinococcosis and alveolar echinococcosis were 1.72% (745/43 362), 1.98% (860/43 362) and 0.02% (8/43 362), respectively. The prevalence of human echinococcosis was higher in pastoral areas (4.13%, 1 577/38 149) than in semi-agricultural and semi-pastoral areas (0.69%, 36/5 213) (χ2 = 151.82, P < 0.05); however, there was no significant difference in the prevalence between men (3.76%, 819/21 787) and women (3.68%, 794/21 575) (χ2 = 0.19, P > 0.05). Cystic echinococcosis was the predominant type in students with echinococcosis (93.78%, 422/450), while alveolar echinococcosis was the predominant type in herders with echinococcosis (72.16%, 801/1 110). There was a significant difference in the constituent ratio of echinococcosis types between students and herders (χ2 = 588.57, P < 0.05). In addition, higher prevalence of echinococcosis was detected in nomadic populations (4.58%, 1 008/22 021) than in community-dwelling populations (2.83%, 605/21 341) (χ2 = 91.88, P < 0.05). The Echinococcus copro-antigen-positive rate was 0.19% (4/2 157) in owned dogs, and the detection rate of echinococcosis was 8.00% (16/200) in yaks and 3.10% (31/1 000) in rodents in 2019. Echinococcosis is highly prevalent in Serthar County, Sichuan Province. Sustained management of source of Echinococcus infections, improved treatment of echinococcosis patients and timely health education for nomadic populations are recommended.

A nationwide study of imported Plasmodium ovale and mixed infections in Israel 2008-2020.

Plasmodium ovale is a rather neglected plasmodium. Rarity, a milder disease, and diagnostic difficulties compared with P. falciparum and P. vivax have led to this situation. This study's objective is to present the epidemiological and diagnostical characteristics of imported P. ovale malaria in Israel. Malaria is a reportable disease in Israel. All highly suspected cases are sent to the Ministry of Health central parasitology laboratory for molecular verification. We retrieved epidemiological and diagnostic data on all polymerase chain reaction (PCR)-proven P. ovale infections imported to between 2008 and 2020. In total, 508 malaria cases were identified, 489 monoinfections and 19 (3.7%) mixed. Fifty-one (10%) were due to P. ovale, among them 13 (25%) were mixed, comprising 68% of all mixed infections. Forty-eight of 51 (94%) underwent blood microscopy, with a sensitivity of 94% (45/48) for genus identification and 15% (7/48) for P. ovale identification. Only 8% (1/12) of mixed infections were identified as such by microscopy. Forty-two (82%) patients underwent RDTs, with a sensitivity of 69% (29/42) for genus identification, and 62% (26/42) for identifying non-falciparum infections. Cycle threshold (Ct) values of P. ovale were significantly higher compared with P. falciparum and P. vivax in both mono and mixed infections (P< 0.05, P< 0.005). Ct levels in RDT positive vs negative infections differed significantly (P< 0.05). P. ovale is commonly imported to Israel from Africa, with a high rate of mixed infections. The use of RDTs and microscopy is insufficient for the species-specific diagnosis of P. ovale, and must be complemented by PCR.

Analysis of comprehensive surveillance results of respiratory tract infectious diseases in Taizhou, 2012-2019

Objective To understand the epidemiological characteristics of influenza-like illness (ILI) cases and severe acute respiratory infection (SARI) cases in Taizhou, so as to provide a basis for scientific prevention and control strategies. Methods The basic information of ILI and SARI cases and throat swab specimens were collected from the sentinel hospital and 13 respiratory pathogens were detected by RT-PCR method.χ2 test was used as statistical analysis method. Results From April 2012 to December 2019, with epidemic peaks in autumn and winter, the percentages of ILI visits (ILI%) and SARI visits (SARI%) were 7.27% and 2.44%, respectively. Finally, 956 ILI case specimens and 557 SARI case specimens were detected. The positive detection rates of the two types of cases were 50.84% (486/956) and 39.14% (218/557), separately. Streptococcus pneumoniae (SP), influenza virus (Flu), Haemophilus influenzae (HI) and Human rhinovirus (HRV) were the dominant pathogens in Taizhou city from April 2012 to December 2019, and their detection rates were 14.81%, 9.25%, 9.39% and 7.73%, respectively. The total mixed infection rate was 30.11% (212/704), which was mainly caused by SP complicated with other pathogenic infections. The positive rate of pathogen detection in the low-age group was higher than that in the high-age group, and the detection rate of children under 5 years old was 34.83% (χ2=54.47, P Conclusions The incidence of comprehensive respiratory tract surveillance in sentinel hospitals in Taizhou City showed that the peak of the epidemic was in autumn and winter, and SP, Flu, HI, and HRV were main pathogens in this area. In the age group under 15 years old, the detection rate was higher. Comprehensive respiratory disease prevention and control measures should be taken for key populations.

Prevalence of malaria and quantification of cytokine levels during infection in East Nile locality, Khartoum State: a cross-sectional study.

Background: The cytokines interferon gamma (IFN-γ), interleukin-10 (IL-10) and tumor necrosis factor-α (TNF- α) play an important role in malaria infection. The aim of this study was to determine the prevalence of malaria and to evaluate cytokine responses to malaria infection in patients from the East Nile locality of Khartoum State. Methods: This study was carried out from May to July 2018 in the East Nile Locality, Khartoum State. Blood samples were collected from 384 randomly selected patients for blood film analysis. Of these, 39 were selected for cytokine level analysis (10 control and 29 patient samples), determined using enzyme-linked immunosorbent assays. Results: The malaria prevalence rate among 384 patients was 18.5%. Plasmodium falciparum was the most prevalent (13%), while the prevalence of Plasmodium vivax was 4.6%. The rate of mixed infection was 0.8%. There was a higher prevalence rate (22.7%) in males than females (15.6%). However, we found no significant correlation between cytokine levels and parasitemia in the study group. Nevertheless, our study demonstrated a significant correlation between cytokine levels and recurrent infections. Conclusions: Together, our data show that malaria remains a public health problem in East Nile locality with a high prevalence. Additionally,serum levels of IFN- γ and TNF- α were significantly higher in malaria-infected individuals compared to non-infecting individuals and cytokine levels were found to be correlated with recurrent malaria infection.