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Overall Solution for Monkeypox Virus Detection of YEASEN

The first confirmed case of monkeypox was reported in the UK in early May, and it quickly broke out in non-endemic countries and regions. As of September 21, 2022, 106 countries/regions have been confirmed worldwide, with a total of 63,117 cases. On September 16, Chongqing confirmed an imported case of monkeypox, the first case in mainland China, which aroused social concern.

Figure 1. 2022 Monkeypox Outbreak Global Map(From the CDC)

Figure 1. 2022 Monkeypox Outbreak Global Map(From the CDC)

 

1. What is Monkeypox?
2. The History of Monkeypox
3. How It Spreads
4. Signs and Symptoms
5. Detection Method
6. Product Match
7. Product Ordering

 

1. What is Monkeypox?

Monkeypox virus is a double-stranded DNA virus with a total length of about 197kb. It belongs to the genus Orthopoxvirus of the Poxviridae family. Virus). There are two branches of the monkeypox virus (West African branch and Congo Basin branch). Among them, the case fatality rate of the West African branch is about 3.6%, and the case fatality rate of the Congo Basin branch can reach 10.6%. Samples of monkeypox cases found in non-endemic areas since May 2022, sequencing analysis showed the West African clade[1].

Figure 2. Monkeypox picture (picture from CCDC)

Figure 2. Monkeypox picture (picture from CCDC)

 

2. The History of Monkeypox

Monkeypox was discovered in 1958 when two outbreaks of a pox-like disease occurred in colonies of monkeys kept for research. Despite being named “monkeypox,” the source of the disease remains unknown. However, African rodents and non-human primates (like monkeys) might harbor the virus and infect people[2].
The first human case of monkeypox was recorded in 1970. Before the 2022 outbreak, monkeypox had been reported in people in several central and western African countries. Previously, almost all monkeypox cases in people outside of Africa were linked to international travel to countries where the disease commonly occurs or through imported animals. These cases occurred on multiple continents[2].

 

3. How It Spreads

It can spread from animals to humans, from humans to humans, and from the environment to humans. 
It is now clear that monkeypox can infect humans in 3 ways: One is from infected animals to people. The virus may enter the body through broken skin, respiratory tract, or mucous membranes (eyes, nose, or mouth). The second is transmitted through close contact between people, including sexual intercourse, kissing, hugging, or touching the body parts of monkeypox, and contact with virus-contaminated clothing, bedding, towels or tableware, and other items that can also be infected. The third is the transmission of respiratory droplets[3].

 

4. Signs and Symptoms

The main clinical manifestations are fever, rash, and lymphadenopathy.
The most common symptoms in the early stage include fever, headache, muscle aches, and swollen lymph nodes. The rash appears 1 to 3 days after the onset of the disease. The typical rash first appears on the face and gradually spreads to the limbs and other parts. The rash is mostly centrifugal. , rashes on the face and limbs are more common than on the trunk, rashes can appear on the palms and soles of the feet, and the number of rashes varies from several to thousands; it can also involve the oral mucosa, digestive tract, genitals, conjunctiva, and cornea.
Symptoms generally last 14 to 21 days, and patients usually resolve them on their own. Erythema or hyperpigmentation or even scars may remain after the scabs fall off. It's worth noting that the scab remains contagious until the scab falls off and a new layer of skin forms underneath[3].

 

5. Detection Method

To effectively prevent and control the monkeypox epidemic and prevent the virus from being imported and spread, global IVD companies have responded actively. As a supplier of IVD detection reagent production raw materials and reagent solutions, YEASEN provides a complete solution to help efficiently detect the monkeypox virus. From nucleic acid extraction to reagent components (enzyme reagents and pseudoviruses), and semi-finished kits (primer probes, enzyme reagents, and plasmid controls), YEASEN can supply you stably.

Figure 3. Virus detection method

Figure 3. Virus detection method

5.1 Method Advantage

  • Efficient and convenient operation—a set of kits, including all components, without cumbersome operations, one-step addition
  • High detection sensitivity—Genetically modified antibody method hot-start Taq enzyme, 1 copies/reaction can also be detected
  • Strong reagent stability—Store at 37℃ for 1 week, stable performance after repeated freezing and thawing
  • UDG Pollution Prevention—The dUTP/UDG system is added to the system to effectively prevent the risk of aerosol contamination.
  • Applicable to multiple platforms—Suitable for Bio-Rad CFX96, ABI Q5, and Slan

5.2 Product Match

  • Nucleic acid extraction reagents: MolPure™ Magnetic Swab Viral DNA/RNA Kit(Cat#18300ES)
  • Monkeypox detection reagent: Monkeypox Virus Real Time qPCR Kit(Cat#13862ES/Cat#13863ES)
  • Monkeypox pseudovirus: Monkeypox virus Pseudovirus(Cat#12099ES/Cat#12100ES)

 

6. Products Recommended

6.1 Monkeypox Virus Real Time qPCR Kit(Cat#13862ES)

Product Advantages:
Efficient and convenient operation
High sensitivity and good specificity
Super stability
Good repeatability

6.1.1 Case Show:

  • Super Thermal Stability for 37℃

The MPXV was amplified by placing Cat#13862ESES at 37℃ for 7 days. The results showed that the Ct value and fluorescence value were not significantly affected by 7 days at 37℃, and the thermal stability was strong.

Figure 4. Yeasen(Cat#13862ES)37℃ accelerated 7-day stability verification

Figure 4. Yeasen (Cat#13862ES) 37℃ accelerated 7-day stability verification

  • Super Freeze-Thaw Stability

The MPXV was amplified by freezing and thawing of Cat#13862ES at -20℃ for 10 times, 20 times, and 50 times, respectively. The results showed that freezing and thawing 10 times, 20 times and 50 times at -20℃ had no significant effect on the Ct value and fluorescence value, and the freezing and thawing stability was strong.

Figure 5. Yeasen(Cat#13862ES)-20℃ freeze-thaw stability verification

Figure 5. Yeasen(Cat#13862ES)-20℃ freeze-thaw stability verification

  • Ultra-high sensitivity, 1 Copies/μL can also be detected

Using Yeasen monkeypox product (Cat#13862ESES), 1000 copies/μL, 100 copies/μL, 10 copies/μL, 1 copy/μL, and negative blank (NTC) were used to detect MPXV. It can be seen that a single copy can also be detected (Ct value is around 38), and The 48-well NTC test was repeated, with a stable baseline, negative NTC peaks, and no tailing phenomenon, with high sensitivity and high specificity.

Figure 6. MPXV sensitivity detection

Figure 6. MPXV sensitivity detection

Figure 7. MPXV specificity detection

Figure 7. MPXV specificity detection

  • Reagents are Stable and Reproducible

The Yeasen monkeypox detection kit (Cat#13862ESES) was selected to carry out 10 repeated tests for low, medium, and high concentrations of positive samples. The results showed that the CV of the positive samples in the FAM channel was less than or equal to 2%, and the blank control had no peaking and no tailing phenomenon. , good repeatability.

Figure 8. Repeatability Test

Figure 8. Repeatability Test

6.2 Monkeypox Virus Real Time qPCR Kit (UDG Plus)(Cat#13863ES)

Product Advantages:

⑴ Efficient and convenient operation
⑵ High sensitivity and good specificity
⑶ Super stability
⑷ Good repeatability
⑸ UDG Pollution Prevention

6.2.1 Case Show:

  • Super Thermal Stability for 37℃

The MPXV was amplified by placing Cat#13863ES at 37℃ for 7 days and 14 days, respectively. The results show that after 7 days and 14 days at 37℃, there is no significant effect on the Ct value and fluorescence value, and the thermal stability is strong.

Figure 9. Yeasen(Cat#13863ES)37℃ accelerated 7-day, 14-day stability verification

Figure 9. Yeasen(Cat#13863ES) 37℃ accelerated 7-day, 14-day stability verification

  • Super Freeze-Thaw Stability

Cat#13863ES was frozen and thawed at -20℃ for 50 times to amplify the MPXV. The results showed that freezing and thawing at -20℃ for 50 times had no significant effect on the Ct value and fluorescence value, and the freezing and thawing stability was strong.

Figure 10. Yeasen(Cat#13863ES)-20℃ freeze-thaw stability verification

Figure 10. Yeasen(Cat#13863ES) -20℃ freeze-thaw stability verification

  • High Sensitivity and Specificity

5 copies/μL of MPXV and negative blank (NTC) were tested using Yeasen monkeypox product (Cat#13863ESES). The results showed that the baseline was stable, the detection rate of FAM was 100%, the minimum detection limit was 5 copies/μL, and during the 48-well repeated testing of NTC, the baseline of FAM and VIC channels was stable, the NTC negative peak did not have any tailing phenomenon, and it had high Sensitivity and high specificity.

Figure 11.MPXV Sensitivity detection         

 Figure 11.MPXV Sensitivity detection

     Figure 12. MPXV specificity detection

Figure 12. MPXV specificity detection

  • Good linearity of Reagents

The positive template was diluted 10 times from the concentration of 1×105 copies/μL, and the Yeasen qPCR monkeypox kit (Cat#13863ES) was used to repeat the detection twice. The logarithm of the initial copy number is the abscissa, and the Ct value is the ordinate. The standard curve was made, and the results showed that the FAM channel of the monkeypox gene had a linear range of 10~1×105 copies/μL, and R2>0.99, which had a good linear relationship.

Figure 13. Linearity Test

Figure 13. Linearity Test

  • Stable and Reproducible Reagents

The Yeasen qPCR monkeypox kit (Cat#13863ES) was selected to carry out 10 repeated tests for low, medium, and high concentrations of positive, and 20 repeated tests for negative control (NC) and blank control (NTC). The results show that: FAM channel The CV of positive samples is less than 3%, and the negative control and blank control have no peaking and no tailing phenomenon; the Ct values of VIC channel NC are all less than 35, CV is less than 0.3%, and NTC has no peaking and no tailing phenomenon, and the repeatability is good.

Figure 14. Repeatability Test(FAM)                   

Figure 14. Repeatability Test(FAM)

   Figure 15. Repeatability Test(VIC)

Figure 15. Repeatability Test(VIC)

 

7. Product Ordering

Yeasen Biotechnology (Shanghai) Co., Ltd., founded in 2014, is a high-tech enterprise engaged in the R &D and production of tool enzyme raw materials and antigen antibodies. Its products include molecular diagnostic enzymes, proteins, and antibodies used in pharmaceuticals, food safety testing, breeding, justice, and other industries. We are committed to providing customers in the field of life sciences with high-quality products and services. The product purchase guidelines for Monkeypox Virus are as follows:

Table 1. Related products

Product Type

Product Description

Product Name

Article Number

Specification

KIT

Enzyme Mix

Monkeypox Virus Real Time qPCR Kit

13862ES03

24T

13862ES48

48T

13862ES59

96T

Monkeypox Virus Real Time qPCR Kit

13863ES25

25T

13863ES60

100T

13863ES80

1000T

Pseudovirus

F3L Gene

Monkeypox virus F3L-Pseudovirus (inquire)

12099ES03

1 mL

12099ES08

5 × 1 mL

B6R Gene

Monkeypox virus B6R-Pseudovirus (inquire)

12100ES03

1 mL

12100ES08

5 × 1 mL

 

References

[1] Chinese Center for Disease Control and Prevention: Monkeypox Health Tips
[2] CDC:About Monkeypox | Monkeypox | Poxvirus | CDC
[3] Yulin Evening News: The source of the first monkeypox case in mainland China has been identified! Guangxi Disease Control Emergency Reminder